During the growth at a few mbar, there is no external carbon gas precursor introduced. The carbon atoms and/or carbon containing particles as a result of the plasma etching are emitted in the chamber, some of which undergo gas phase scattering and deposit onto the metallic catalyst substrates  as graphene sheets. It is found that high quality monolayer graphene can be synthesized on Cu foil at 900 °C. On Cu-Ni, under the same growth condition, somewhat more bilayer regions are observed. It is observed that the material quality is almost indifferent to the gas ratios, which makes the optimization of the deposition process relatively easy. Detailed study on the deposition procedure and the material characterization have been carried out. This work reveals the possibility of producing thin film graphene by a gas discharge based process, not only from fundamental point of view, but it also provides an alternative technique other than standard chemical vapor deposition to synthesize graphene that is compatible with the semiconductor planar process. As the process uses solid graphite as a source material that is rich in the crust, it is a facile and relatively cheap method to obtain high quality graphene thin films in this respect.Arc discharge is traditionally used to synthesize randomly arranged graphene flakes. In this paper, we substantially modify it into a glow discharge method so that the discharge current is much more reduced. The H Since its appearance, graphene has been applied to solar cells, sensors, composite materials, photocatalysis, and other fields due to its outstanding characteristics such as high carrier mobility, high mechanical strength, high specific surface area, high transmittance, and high thermal conductivity . The gra2 and/or Ar gas, we ignited the plasma, and it slightly etches the graphite electrode that is used to start the plasma. Because of this, we were able to grow graphene on the surface of metal catalysts even without using any carbon precursor gas. The possible deposition mechanism is explained by the plasma-based physical and/or chemical etching of the graphitic electrode, followed by gas phase scattering of the produced carbon atoms and carbon based particles, which are in part transported towards the catalytic surfaces of the metallic substrates situated on the heater. After the catalytic graphitization on the metal surface, the prepared graphene/metal is unloaded, and the graphene can be transferred to insulators using a wet etching-based technique [In this paper, we have substantially modified the traditional AD method, based on which we introduce a new graphene synthesis technique that is called the glow discharge (GD) deposition method. The equipment used in this work is actually a standard PECVD (plasma enhanced CVD) system that was not originally intended for the GD use. In the chamber, which is filled with Hechnique ,15. The In order to verify and benchmark the quality of the graphene obtained by the new GD graphene production mode proposed in this paper, we first grow standard graphene thin films by CVD using two kinds of conventional catalytic metals  ,17 as co2 is grown by inductively coupled plasma chemical vapor deposition (ICP-CVD). This is because, later, the Cu-Ni will be removed and the grown graphene will “land” on the SiO2, and 300 nm silicon dioxide is known to facilitate the observation of graphene under an optical microscope after growth, due to the optical interference effect [2/Si wafers to study the effect of catalyst thickness on the quality of graphene. (1) Cu-Ni alloy thin films. After the Si wafer is cleaned by standard procedure, 300 nm SiOe effect . Magnetr(2) Copper foils. High purity polycrystalline copper foils are purchased commercially.The growth system used in this work is a graphene PECVD of the model Black Magic, produced by Aixtron Nanoinstruments Ltd. The outwall of the growth chamber is made of stainless steel with a quartz shield as the inwall. As shown in −3 kPa, we introduce Ar and/or H2 gases  into the chamber, and the pressure reaches 5 × 10−1 kPa. The procedure is repeated three times and then the gas input is turned off. This is because we have found that without a flowing gas, the plasma power during the initiation is more stable than is the case with a gas flow. The heating rate is 5 °C/s, at which we increase the heater temperature to the desired growth temperature . The samples are held at this temperature for ten minutes for annealing to increase the crystallinity of the metals [We place the as-prepared Cu-Ni alloy thin film or copper foil samples on the graphite plate heater in the equipment see  for the e metals , which i4:HCl:H2O = 5 g:25 mL:50 mL) is prepared. The copper-nickel sample is placed at the bottom of the beaker, whereas the copper foil sample is floating on the surface of the solution. The copper foil grown graphene is transferred to another 300 nm SiO2/Si substrate by the standard wet transfer process [2/Si substrate through the etching mechanism that is shown in 2/Si complex in deionized water for 30 min to clean up the residual chemicals. In order to improve the adhesion between the graphene and the substrate, it is baked at 150 °C for 15 min on a hot plate. Then, it is placed in acetone for 1 h to remove the PMMA from graphene. Finally, the sample is placed in a ventilated place for 10 min to dry the acetone. During the experiment, nevertheless, after the metal etching the van der Waals force between the SiO2/Si substrate and the PMMA/graphene is relatively weak, and the rinsing process in deionized water might cause the PMMA/graphene to float on the surface of solution. Applying a little PMMA to the edges of the sample helps hold the film, and can increase the success rate of the graphene transfer.After growth, the copper-nickel alloy and the copper foil samples are all spin-coated (4000 rpm for 30 s) with a layer of polymethyl methacrylate (PMMA) with about 70 nm thickness. Then, it is heated on a 150 °C hot plate for 10 min. A metal etching solution  is used to examine the morphology of the metals after the annealing and after the graphene deposition.2 gas ratios are used, we can always obtain thin film graphene conformally coating on the metal catalysts, provided that other conditions are optimized . Since there is no carbon containing gas introduced in the machine, the only explanation is that the graphene grows from the etched graphite. Note that we have found the graphene thin films do not form when no plasma is present. Thus, we can conclusively exclude the possibility that the graphene’s carbon source comes from unintentionally introduced carbon species in the chamber such as carbon contamination, oil vapor from the pump, etc. In our experiment, the etching mechanisms of H2 and Ar plasmas are different. The former (H2 plasma) is mainly a chemical process, where the etching of graphite is achieved through hydrogenation (forming C-H bonds) and the subsequent releasing of hydrocarbons such as CH4 [λ of the particles can be approximately estimated by the ideal gas formula d is the effective diameter of the particle, and n is the number of particles per unit volume. Using that equation, the mean free path of the carbon-based particles is in the order of a few millimeters, which is much shorter than the distance between the plasma electrode and the samples on the heater (~4 cm). Therefore, the particles undergo many scatterings in the chamber and some particles will have chances to be directed toward the samples, as depicted in The schematic illustration of the graphene deposition procedure is shown in h as CH4 . Some auh as CH4 . The lath as CH4 ,24. So, 2 and/or Ar gases are required in the chamber. As can be seen later, the quality of graphene is insensitive to the H2 to Ar ratio. According to our experiment, the material quality can be optimized by adjusting parameters such as the type of catalytic metals and the deposition temperature. Finally, after the GD growth, we have measured the 300 μm thick plasma electrode and could not detect any weight loss. Therefore, the process is indeed very gentle, and the consumption of the graphite material is tiny. In our practice, the same graphite electrode can be used for hundreds of runs.The introduction of the GD production mode in the PECVD chamber makes it feasible to grow thin film graphene under our conditions, which is not possible in the traditional AD method, which can only produce chaotic flakes. That is a new feature in graphene synthesis by gas phase discharge, both from fundamental and application points of view. Furthermore, in the GD method, the graphene is grown from cheap graphite and not an expensive carbon precursor gas, reducing the cost in this regard. Also, because there is no carbon precursor gas, it makes the process optimization much easier, without the need to monitor the precursor gas ratios. Only H2 hybridized graphitic carbon structure. As the temperature rises from 600 °C to 1000 °C, the characteristic 2D peaks of the graphene Raman spectroscopy gradually appear. Meanwhile, the D peak eventually decreases. These features indicate that the number of defects or disorders in the graphene decreases. The Raman characterization results of the samples at 800 °C–1000 °C are more or less similar to each other. The Raman results are also comparable with standard CVD graphene grown in the same type of machine , whi2/Si substrates. It is well established that the number of layers in graphene with good crystallinity can be estimated through the 2D/G ratio, and the shape and width of the peaks [In order to examine the structural uniformity of the graphene along the sample, more detailed characterizations are carried out. he peaks . It can he peaks , becausehe peaks  . Theref2/Si) grown on a 300 nm Cu-Ni alloy with different gas ratios at 900 °C. The gas mixture is varied from pure Ar, Ar:H2 = 5:1, Ar:H2 = 5:3, Ar:H2 = 5:5, to pure H2. It can be seen that the graphene quality almost does not change, which proves that the GD mechanism does not depend much on the Ar-H2 composition of the gas. No matter whether it is a large atom Ar gas or a small molecule H2 gas, the GD can etch the graphite electrode and supply carbon source anyway . Therefore, in future experiments, cheaper gas can be selected to prepare the graphene by this method. However, because the reaction temperature is typically above 800 °C, oxygen or air cannot be selected as the plasma gas so as to avoid the burning of the graphite.2/Si) grown at 900 °C with a different thicknesses (from 20 to 300 nm) of Cu-Ni alloy and an Ar:H2 = 5:1 gas ratio. The original idea of this experiment was to reduce the amount of absorbed carbon (hence reducing the number of produced graphene layers) through reducing the thickness of the Cu-Ni alloy. However, the experimental results show that even if the 20 nm Cu-Ni alloy is used, it still can produce bilayer and multilayer graphene. On the other hand, as the thickness decreases, the D peak rises and more defects appear. This article considers that the graphene quality gets worse at reducing thickness of the alloy because during the growth, the metals of the 20 nm and 50 nm samples sublimate from the surface under high temperature and low pressure conditions, which gradually leads to the loss of the graphene catalysis effect. If we continue to test the growth on even thinner metals, the graphene quality will be even worse, and the alloy will melt due the lowering of the melting point with the reduction of the thickness. Towards the other end, when the thickness of the Cu-Ni alloy reaches 100 nm and above, the quality of graphene has gradually stabilized.2/Si substrate. The ratio of ID to IG is less than 0.35, and the ratio of IG/I2D is mostly around 0.5. This proves that the uniformity of the number of graphene layers is very good across the sample and the graphene is mainly single layer. As indicated earlier, we have also grown graphene on copper foils by the GD method in order to boost the monolayer ratio. The corresponding Raman measurement results are demonstrated in 2 and/or Ar plasma environment, and high-quality graphene thin films can be synthesized at about 900 °C on catalytic metals e.g., Cu-Ni alloy and pure Cu. The graphene production mechanism is a plasma etching of the graphite, followed by a carbon containing particle emission, where the particles are scattered towards the catalytic metal surfaces for graphene thin film formation. To our knowledge, this graphene production mode has never been reported before. It reveals that graphene sheets (not irregular flakes) can be obtained by gaseous discharge-based technology. The as-grown graphene is transferrable, and is characterized by Raman spectroscopy, SEM, and other means to prove the feasibility of the GD. It is found that the major factors during the graphene synthesis that can determine the material quality are the growth temperature and the type of catalytic metals, but not the gas ratios. This work shows that the GD graphene production model can be used to prepare graphene thin films without using carbon-containing gases such as methane and ethylene, which provides a new technology and a new insight for graphene synthesis. In future work, we will continue to improve the quality of graphene and increase the sample size. We will apply the graphene in semiconductor devices, such as current spreading layers of light-emitting diodes [In this paper, we report a new type of graphene synthesis technology called GD deposition, using a standard PECVD machine. Compared to the traditional AD method, the GD technique uses a much smaller current, and the etching process of the graphite electrode that is used to ignite plasma is very gentle. The graphite electrode can be directly used as the carbon source for the graphene growth in a Hg diodes . We willg diodes  or carbo


Two distinct cases we considered: (i) the Au NPs sit with their (111) facets on graphene and (ii) the NPs are oriented with a vertex on graphene. In both cases, we compare the binding energies with and without a graphene vacancy under the NP. We find that in all cases, the presence of the graphene vacancy enhances the bonding of the NPs. Significantly, in the vertex-on-graphene case, the binding energy is considerably increased by several eVs and becomes similar to the (111) facet-on-graphene case. The strain in the NPs is found to be minimal and the displacement of the carbon atoms in the immediate neighborhood of the vacancy is on the 0.1 Å scale. The work suggests the creation of stable NP-graphene systems for a variety of electronic, chemical and photonic applications.Hybrid nanostructures of size-selected nanoparticles (NPs) and 2D materials exhibit striking physical and chemical properties and are attractive for many technology applications. A major issue for the performance of these applications is device stability. In this work, we investigate the bonding of cuboctahedral, decahedral and icosahedral Au NPs comprising 561 atoms on graphene sheets via 10 The unique physical and chemical properties of 2D materials—graphene being the primary example—and of nanoparticles (NPs) can be further enhanced by combining the two in hybrid nanostructures. Indeed, NP-graphene composites show promise for electrochemical energy applications such as lithium batteries, supercapacitors and catalysts for various reactions including oxygen reduction, oxygen evolution and hydrogen evolution reactions  and a vaThe triangular lattice of the 111) surface of the face-centered cubic (fcc) metals is a good match for the hexagonal honeycomb lattice of graphene, and the metals are accommodated on graphene with minimal strain depending on their lattice constant. An ab initio investigation reported weak bonding between graphene and Al, Ag, Cu, Au and Pt, with a binding energy of 0.03 eV per carbon atom for Au 1 surface. A deepe2700 clusters produced via a magnetron sputtering, gas aggregation cluster beam source were deposited on highly oriented pyrolytic graphite (HOPG) predecorated with surface defects created by Ar+ sputtering. The subsequent scanning tunneling microscopy (STM) investigation revealed a stable, randomly dispersed array of the clusters. The effect of surface modifications on the anchoring of Au NPs on carbon surfaces was showcased in Ref. [The bonding of noble-metal nanoclusters on carbon surfaces has been shown to be enhanced by surface modifications. In Ref. , Ag2700  in Ref.  where sm in Ref.  using mo in Ref.  researchIn this work, we aim to overcome previous theoretical studies in terms of completeness and size. We employ ab initio calculations of a scale not previously reported in the literature with realistic models to examine and quantify from first principles, the effect of surface modifications, specifically vacancy defects, on the bonding, energetics and structure of Au NP-graphene hybrid nanostructures. We attempt a thorough examination of the system and check for three highly ordered NP structural motifs and two different cluster orientations. We take into account both spin-polarization and van der Waals interactions. Our results show that the creation of vacancies strengthens the bonding of the Au NPs on graphene, particularly when the NPs sits with a vertex on the support. The resulting nanostructures show a symmetrical displacement of the C atoms surrounding the vacancy and minimal strain in both the NPs and the graphene sheet. The results of this work can be expanded to other NP sizes and cluster materials and exploited for the creation of stable and efficient NP-graphene devices.The spin-polarized DFT calculations were performed using the Vienna Ab initio Simulation Package  ,19 underWe focus on 561-atom nanoclusters that lie in the catalytically active regime. Such clusters are routinely produced by cluster deposition methods such as the gas-phase condensation method. Choosing a “magic number” for the nanoclusters also allows for a ready comparison between the three highly ordered structural motifs, the cuboctahedral, the Ino-decahedral, and the icosahedral, which are shown to be dominant for Au NPs of this size . We placa lattice constant of the graphene unit cell was found to be 2.468 Å. The choice of the correction method was made based on the results of Ref. [Van der Waals interactions were taken into account through the DFT-D3 method  with Bec of Ref.  on the c of Ref. , the cat of Ref. , the ele of Ref. , and to  of Ref. . The imaging of the results was performed with the VESTA 3.4 visualization program  . The dataset generated during the current study is available online at the Zenodo repository .The (111) facets of the NPs were interfaced with graphene following the energetically favorable configuration of according to Ref. : the atoThe most common and energetically favorable point defect of graphene is the single vacancy. We calculated the formation energy  studies . The C a561 clusters. To this end, we ran a set of relaxations for three distinct locations of the vacancy under the triangular (111) facet of a cuboctahedral NP: (i) vacancy under one edge of the triangle, (ii) vacancy under a vertex of the triangle and (iii) vacancy near the center of the triangle. These positions are shown in After the initial set of relaxations of the three isomers on pristine graphene, we investigated the energetically preferred position of the vacancy under the (111) facet of the AuThe binding energies In pristine graphene, all C atoms present the same charge with only minimal changes in the scale of 0.01 In the case where the cluster vertex points at the graphene substrate, we initially investigated the bonding site of the NPs with a vertex on pristine graphene and ran relaxations for two distinct positions: (i) the vertex of the NP sits on top of a C atom and (ii) the vertex of the NP lies in the center of the C-atom hexagon . We founWe explored two different symmetrical orientations for each NP isomer with a vertex on a vacancy, as shown in The corresponding binding energy results after relaxation are shown in The addition of the NP at the vacancy site was found to remove the Jahn-Teller distortion in the C atoms surrounding the vacancy. An example of a vertex-on-graphene case is shown in The charge density difference Where The electronic properties of graphene are affected by the presence of the NPs. In Finally, the energetic ordering of the NPs is unchanged by the interaction with the graphene sheet. The stability of the bare NPs follows the icosahedral > decahedral > cuboctahedral trend reported previously in previous theoretical studies of regular clusters in the literature , with an3 atoms in ab initio calculations that took into account spin-polarisation and van der Waals interactions. Our results showed that defects enhance the bonding of Au561 clusters with three atomic structures on graphene, particularly when the clusters sit with a vertex on the vacancy. The resulting binding energies in the last case were similar to the facet-on-graphene case, despite the involvement of only 1 Au atom in the bond with graphene, compared with 21 in the parallel-facet case. All of the resulting nanostructures showed a small strain, a symmetrical displacement of the C atoms in the immediate neighborhood of the vacancy, a uniform distribution of the charge between these C atoms, and a minimal strain on the NPs. The increased binding energy and the competition between facet-down and vertex-down are expected to be even more pronounced in clusters smaller than 561 atoms, since fewer atoms belong to the (111) facets. In contrast, the number of atoms in the vertex is independent of the NP size. We generally expect the vertex-on-graphene binding energies to remain constant with the size of the NPs, and the facet-on-graphene binding energies to increase with the size of the NPs.We investigated the energetics and the electronic, magnetic, and structural effects of graphene vacancies on Au NP-graphene hybrid nanostructures in this work. To do so we used large-scale models consisting of more than 10We believe that a closer look into the electronic properties of these nanostructures would be of great interest for future studies and critical for their application in electronic devices. The methodology of this paper can also be applied for the examination of the properties of another novel hybrid nanostructure of NPs and graphene: graphene-covered gold NPs ,38. Such


Idiopathic pulmonary fibrosis (IPF) is associated with an increased risk for lung cancer, but the underlying mechanisms driving malignant transformation remain largely unknown. This study aimed to identify differentially expressed genes (DEGs) distinguishing IPF and lung cancer from healthy individuals and common genes driving the transformation from healthy to IPF and lung cancer.The gene expression data for IPF and non-small cell lung cancer (NSCLC) were retrieved from the Gene Expression Omnibus (GEO) database. The DEG signatures were identified via unsupervised two-way clustering (TWC) analysis, supervised support vector machine analysis, dimensional reduction, and mutual exclusivity analysis. Gene enrichment and pathway analyses were performed to identify common signaling pathways. The most significant signature genes in common among IPF and lung cancer were further verified by immunohistochemistry.MMP-1) in this pathway was mutually exclusive with several genes in IPF and NSCLC. Subsequent immunohistochemical analysis verified enhanced MMP1 expression in NSCLC associated with IPF.The gene expression data from GSE24206 and GSE18842 were merged into a super array dataset comprising 86 patients with lung disorders (17 IPF and 46 NSCLC) and 51 healthy controls and measuring 23,494 unique genes. Seventy-nine signature DEGs were found among IPF and NSCLC. The peroxisome proliferator-activated receptor (PPAR) signaling pathway was the most enriched pathway associated with lung disorders, and matrix metalloproteinase-1 (For the first time, we defined common signature genes for IPF and NSCLC. The mutually exclusive sets of genes were potential drivers for IPF and NSCLC. Idiopathic pulmonary fibrosis (IPF) is a chronic, progressive, and usually fatal interstitial lung disease that is characterized by dysfunction and damage of lung epithelial cells and aberrant pulmonary remodeling. After diagnosis, patients usually have a median survival of 3–5 years, and the main cause of death is respiratory failure , 2. AlthLung cancer is the most common malignant tumor and the leading cause of cancer deaths worldwide . Non-smaIn this context, we aimed to identify common genes involved in both IPF and lung cancer by screening the gene expression omnibus (GEO) database for gene expression profile data of lung tissue samples from healthy controls as well as from patients with IPF and NSCLC. A multistep strategy Fig.  was applhttp://www.ncbi.nlm.nih.gov/projects/geo/) [Homo sapiens. In this study, the gene expression datasets obtained from the Affymetrix Human Genome U133 Plus 2.0 Array were used as the discovery cohort for data modeling, and the expression datasets obtained from other assays served as the validation cohort. After recognizing the raw data from the IPF and NSCLC patients as well as the healthy controls, probes of the array data were replaced with the corresponding official gene symbols using the GPL570 platform. Duplicates were collapsed using probe medians, as described previously [The gene expression profile datasets on IPF and NSCLC were searched and downloaded from the GEO repository (ts/geo/) . The keyeviously , 15. Logeviously . During eviously , 18.P < 0.05 were considered as DEGs. During the TWC process, hierarchical clustering algorithms based on the Euclidean distance as a similarity metric and a complete linkage clustering approach were implemented. The hierarchical clustering analysis and visualization were performed using Cluster 3.0 and Java Treeview software. The normalized gene expression data were matched with the DEGs identified by the Student’s t-test to generate the filtered gene expression data with the DEGs, which were subjected to supervised classification via support vector machine (SVM) analysis using multtest and kernlab in the R software package [The unpaired Student’s t-test and two-way clustering (TWC) analysis were used to identify genes that were expressed differentially between disease (IPF and NSCLC) and control samples; genes with  package  and vali package . The SVM package . The LOOP < 0.05 were defined as significantly altered. Each principal component in each gene has a corresponding loading coefficient. Genes with a loading coefficient with an absolute value > 0.6 were selected as signature genes.Microarray data possess high-dimensional properties. Each gene may have interactions with others, and any potential interactive relationship is within the data. Principal component analysis (PCA) was utilized to extract key components from the high-dimensional gene expression data. The filtered data with DEGs that were identified by supervised classification analysis (weight coefficient > 1.0) were analyzed with PCA for dimensionality reduction using the prcomp function in the R software. Briefly, the principal components of each DEG were extracted, and the loading coefficients of each gene were calculated, which can be used to assess the ability of the gene to promote or inhibit a disorder (IPF/NSCLC). The standard deviation, proportion of variance, and cumulative proportion are shown in detail in the To validate the models independently, the GSE10667 dataset , which whttps://david.ncifcrf.gov/) [https://www.kegg.jp/) were used to annotate the functions and pathways as well as gene enrichments of the signature genes that were differentially expressed between IPF/NSCLC patients and healthy controls.The Database for Annotation, Visualization, and Integrated Discovery  , 25 and k  and t (number of modules) values were determined using the criteria outlined previously [t = 2 and k = 2 were selected for marginal probability modeling . Gene pairs within the mutually exclusive module likely drive disease progression and thus were defined as putative cancer genes [Synthetic lethality is a type of genetic interaction in which the simultaneous functional loss of two or more genes through mutations, amplifications, or deletions leads to cell death. This genetic phenomenon is vital for cell viability and is emerging as a novel therapeutic target for cancer treatment . Pairs oeviously ; and as er genes . Common To validate the signature genes identified by gene expression modeling, the expression pattern of the signature genes showing mutual exclusivity was analyzed in patients diagnosed with IPF or lung cancer. The Ethics Committee of Beijing Chao-Yang Hospital approved the study and waived informed consent (2017-Science-10). Paraffin-embedded lung tissue specimens were obtained from the archives in our hospital from patients with IPF, NSCLC without IPF, and NSCLC associated with IPF for immunohistochemical (IHC) analysis. In addition, tissue specimens obtained from healthy donor or adjacent normal tissue of NSCLC were used as the reference. Table MMP-1)  at 4 °C overnight, and then incubated with the secondary antibody for 20 min at room temperature. Finally, diaminobenzidine (DAB: ZSGB-BIO ZLI-9017) staining was performed.After deparaffinization with methanol and ethanol and placement in ethylenediaminetetraacetic acid (pH 8.0) for antigen retrieval, the retrieved paraffin-embedded tissue sections were incubated with the polyclonal primary antibody against matrix metalloproteinase-1  and 51 healthy control lung tissue samples.P < 0.05  and the number of signature genes, which were defined as an absolute loading coefficient > 0.6, for all comparisons. As illustrated in Fig. PCA was conducted to identify the most significant genes that distinguish healthy subjects from patients with lung disorders. The first principal component (PC1) was found to be the most significant in all comparisons of healthy controls with patients with IPF and NSCLC . KEGG pathway analysis showed that the common signature genes were closely related to the peroxisome proliferator-activated receptor (PPAR) signaling pathway and metabolic pathways (Table lipoprotein lipase (LPL), and MMP1 were the four common signature genes in the PPAR signaling pathway; and ACADL, amine oxidase, copper containing 3 (AOC3), hydroxysteroid 17-beta dehydrogenase 6 (HSD17B6), and ribonucleotide reductase family member 2 (RRM2) were in the metabolic pathways. In addition, three common signature genes in the pathways concerning cancer were endothelin receptor B (EDNRB), hedgehog-interacting protein (HHIP), and MMP1.The 79 common signature genes were uploaded to DAVID and KEGG for gene enrichment and pathway analysis. Table ys Table . The PPAThe MCMC method was used to identify mutually exclusive modules in the comparison of healthy controls with IPF and NSCLC. When the edge weight was set to δ = 0.2, three different types of cliques were generated for these comparisons; while when δ = 0.1, more mutually exclusive modules were obtained for each comparison (data not shown). Seven cliques involving 33 genes and 11 cliques involving 39 genes were generated in the comparisons of healthy controls with IPF and NSCLC, respectively. Among these genes, MMP1, PPAP2C, SFTA1P, and LPL were found to be shared between IPF and NSCLC. MMP1 was mutually exclusive in the two comparisons and had a high coverage power Fig. . MMP1 waThe gene expression of MMP1 was further validated in lung tissue samples from IPF patients, NSCLC patients without IPF, and NSCLC patients with IPF by IHC staining. Positive IHC staining of MMP1 was observed in all of the tested samples, but the staining intensities were different. A slightly increased expression of MMP1 was observed in the NSCLC-without-IPF tumor tissue and the IPF tissue compared with the adjacent normal lung tissue sections or with the healthy donor lung tissue. MMP1 expression was mostly observed in the alveolar epithelium in the IPF tissue; whereas in the NSCLC-without-IPF tumor tissue, MMP1 presented predominantly in the glandular epithelium. The NSCLC-with-IPF tumor tissue displayed a distinctly stronger expression of MMP1 compared with the other tissue samples, and the staining was mainly localized to the glandular epithelium and extracellular stroma. Specifically, MMP1 staining was more intense in the tumor section of NSCLC with IPF compared with the paired IPF section, and compared with the tumor section of NSCLC without IPF. Particularly, there was no association between MMP1 expression and tumor staging, but MMP1 expression was probably correlated with the severity of IPF Fig. .Fig. 7IPrevious studies have indicated that patients with IPF are at high risk for lung cancer , 31, 32,In this study, we examined the gene expression profile data of IPF and NSCLC from the GEO repository using bioinformatic analysis. MMP1 was identified as a promising cancer driver gene related to the transformation from healthy cells to IPF, and from healthy cells to NSCLC. MMP1 expression was further validated in IPF and NSCLC tissues by IHC, and the results could partly confirm the enhanced expression of MMP1 in NSCLC associated with IPF. Studies with more tissue samples are needed to quantitatively confirm these findings. Interestingly, there were no differentially expressed genes between IPF and NSCLC samples (data not shown). This result partially confirms the well-recognized opinion that from the perspective of genetic alteration, IPF shares common hallmarks in response to fundamental pathogenic events like cell proliferation, myofibroblast origin, etc. , 37. To In this study, the gene expression profile data from IPF and NSCLC patients as well as healthy controls were pooled and clustered to visualize the similarities and differences in their expression. The gene expression patterns associated with these disorders were gradually filtered through unsupervised and supervised classification and dimensional reduction; distinct expression patterns were observed. The SVM classifier and PCA used in this study are novel tools for biological data mining and have been successfully applied previously for lung disorder classification , 41. TheIt is reasonable to hypothesize that the common signature genes shared by IPF and lung cancer are gene signatures for lung cancer associated with IPF and that signaling pathways enriched by these genes may be involved in disease progression. In this study, the most enriched pathway was the PPAR signaling pathway, which is involved in cell proliferation. Although the exact mechanism of PPARs in lung fibrosis and lung cancer is largely unknown, a PPARγ agonist has been found to exert antitumorigenic effects in both IPF and lung cancer by inhibiting myofibroblast differentiation and activating phosphatase and tensin homolog (PTEN) , 43, conIn this study, we identified four common cancer genes that were shared in common between IPF and NSCLC and were mutually exclusive, while each of these genes had their own mutually exclusive partners for each lung disorder. It is generally accepted that genetic alterations that drive cancer are often mutually exclusive . TherefoThe outliers observed during the analysis were probably due in part to the epigenetic influence of DNA methylation, histone tail modification, and noncoding RNAs, as suggested previously, which are influenced by other diseases, aging, smoking, diet, and other environmental stimuli . BecauseTo the best of our knowledge, this is the first study to compare the gene expression profiles across healthy, IPF, and NSCLC samples using bioinformatic analysis based on published gene expression data. We identified signature genes common to IPF and lung cancer as well as the common signaling pathways involved in tumor development. By introducing mutually exclusive expression analysis, we found potential driving modules for both IPF and NSCLC. Of these genes, MMP1 appeared to be the most promising driver gene showing significant differential expression in the transformation from healthy to IPF and NSCLC, supporting its potential as a novel therapeutic target for IPF, lung cancer, and IPF-progressed lung cancer. Further investigations to verify the gene expression patterns identified in this study in more samples with quantitative experiments and to explore the underlying mechanism in primary cells from IPF, lung cancer, and IPF-associated lung cancer are needed to provide new insight into lung tumorigenesis and targeted therapy.Additional file 1. PCA summaries for Healthy vs. Disorder, Healthy vs. IPF, and Healthy vs. NSCLC. Bayesian probit regression results for Healthy vs. Disorder, Healthy vs. IPF, and Healthy vs. NSCLC.


The diverse structural and electronic properties of the Si-adsorbed and -substituted monolayer graphene systems are studied by a complete theoretical framework under the first-principles calculations, including the adatom-diversified geometric structures, the Si- and C-dominated energy bands, the spatial charge densities, variations in the spatial charge densities and the atom- and orbital-projected density of states (DOSs). These critical physical quantities are unified together to display a distinct physical and chemical picture in the studying systems. Under the Si-adsorption and Si-substitution effects, the planar geometric structures are still remained mainly owing to the very strong C–C and Si–C bonds on the honeycomb lattices, respectively. The Si-adsorption cases can create free carriers, while the finite- or zero-gap semiconducting behaviors are revealed in various Si-substitution configurations. The developed theoretical framework can be fully generalized to other emergent layered materials. The Si-doped graphene systems might be a highly promising anode material in the lithium-ion battery owing to its rich potential properties. The versatile morphologies directly indicate the peculiar chemical bondings, in which all carbon-created systems possess 16 since the first experimental observation in 2004 by mechanical exfoliation. Up to now, they clearly exhibit plenty of remarkable fundamental properties due to the hexagonal symmetry, the nanoscaled thickness, and the distinct stacking configurations, such as semiconducting and semi-metallic behaviors18, anomalous quantum Hall effects19, diverse magnetic quantizations23, rich Coulomb excitations and decays28, different magneto-optical selection rules31, the exceedingly high mobility of charge carriers33, and the largest Young’s modulus of materials ever tested34. To induce the novel phenomena and extend the potential applications, the electronic properties can be easily modulated by the layer number36, stacking configuration39, mechanical strain41, sliding42, electric and magnetic field44, atom adsorption48 and substitution51. This paper mainly focuses on the latter two factors.Carbon atoms can form three-dimensional (3D) diamond53, molecules54 and even functional groups55. To date, many theoretical56 and experimental research papers on surface adsorptions have been published57. The previous results show adsorption-diversified physical and chemical phenomena, such as an opening of the energy gap58, semiconductor-metal transitions62, the absence/recovery of a Dirac-cone structure63, spin-splitting energy bands due to specific adatoms64, single- or multi-orbital hybridizations in C-adatom bonds shown in the spatial charge density and atom- and orbital-decomposed DOSs65, and the shift of the Dirac cone of graphene field effect transistors (GFETs) under doping with the polyethylenimine (PEI) molecule66. It is well known that graphitic systems are the most efficient anode material in Li67 and put to use for nano-scaled applications. Another chemical modification is the direct doping, the substitution of carbon host atoms by guest adatoms in the honeycomb lattice. For example, diversifying the physical and chemical properties of graphene by the silicon doping effects68, modifying the properties of graphene by exactly positioning atomic dopants of nitrogen or phosphorus70. Besides, the B71 and 2D layers72. Apparently, the essential properties are predicted to present drastic changes under the adatom dopings, e.g., in the electronic structures73, optical absorption spectra74 and transport properties75. The feature-rich properties of Si-adsorbed and Si-substituted graphene systems are worthy of a complete systematic study, especially for the critical differences in the orbital hybridizations between the weak and strong Si–C bonds.How to modulate the fundamental properties becomes one of the main-stream topics in materials science, chemistry, physics and engineering. The chemical modification is the most effective method. Pristine graphene has a rather strong In this paper, a complete theoretical framework, which is developed by the first-principles calculations, is utilized to fully explore the feature-rich properties of the Si-adsorbed and Si-substituted graphene systems. The concise chemical and physical pictures and the multi-orbital hybridizations will be proposed to explain the adatom diversified geometric structures, electronic energy spectra, spatial charge densities, and atom- and orbital-decomposed density of states. That is, all the calculated results are consistent with one another under such mechanisms. Furthermore, they are responsible for the semiconductor-metal transitions after the guest-atom adsorptions and the creation of energy gaps in the substitution cases. The theoretical predictions can be thoroughly verified by the various experimental measurements, including the scanning tunneling microscopy (STM)/transmission electron microscopy (TEM), angle-resolved photoemission spectroscopy (ARPES) and scanning tunneling spectroscopy (STS).76, germanene77, and tinene78. Apparently, this crystal is formed by the very strong Monolayer graphene has a planar geometry with a honeycomb lattice, being different from the buckled structures in layered silicenementclass2pt{minimmentclass2pt{minimmentclass2pt{minimmentclass2pt{minim79, Hall effects80 and optical properties81, being consistent with the experimental measurements82. The linear energy dispersions will gradually become parabolic ones as the state energy of A diagram of the first Brillouin zones for various unit cells is presented in Fig. The electronic band structure of the pristine systems is fully changed under the Si adsorptions. The band asymmetry about the Fermi level becomes more obvious. For the The atom- and orbital-dominated energy bands are worthy of a closer examination. For 100Both silicon and alkali adatoms can create free carriers, while their band properties are quite different from each other. The alkali-adsorbed graphene systems present approximately rigid energy bands and a few Li-dominated conduction bands, clearly indicating the blue shift of the Fermi level. Their free carriers purely originate from the electron charge transfer from the outermost s orbital of each alkali adatom to the carbon host atom. Furthermore, the atom- and orbital-projected densities of states clearly show the weak, but significant s-pElectronic band structures obviously change in the presence of Si substitutions, as clearly indicated in Fig. z-direction (area covered by a red rectangle). The 2px-, y-, and z-directions present obvious variations. The strong evidences are illustrated by z, x)- and -planes (red regions). This result means that some electronic charges transferred from Si adatoms to C atoms. In addition to the z-direction, the important charge variations along the x- and The multi-orbital hybridizations in chemical bonds, which are responsible for the adatom-diversified geometric structures, electronic band structures and density of states, can be delicately identified from the spatial charge densities - and -planes, being clearly show in Fig. The Si-substitution cases exhibit diversified charge densities compared with the pristine and Si-adsorption ones. For the 100ice Fig. a, presenice Fig. a. They ads Fig.  a. This ions Fig. c,e, and ene Fig. a. The obThe main features of electronic band structures can be fully identified in the density of states. Special structures, the van Hove singularities, mainly originate from the critical points in the energy-wave-vector space, in which the band-edge states might belong to the local minima and maxima and the saddle points. In general, there are three kinds of novel structures, the V-shaped structure crossing at the Fermi level, logarithmically divergent peaks [Apparently, a lot of van Hove singularities in the density of states are created by the Si adsorption in the ons Fig. b–g. HoweThe substitution and adsorption cases are thoroughly different from each other in the main feature of density of states. For the former chemical modifications, the number of electronic states, which is revealed in Fig. 84, 1D metallic and semiconducting carbon nanotubes86, and 3D Bernal graphite87. Apparently, the theoretical predictions on the Si-adsorption- and Si-substitution-diversified density of states in monolayer graphene systems, which could be examined by the STS experiments, cover the finite or vanishing density of states at the Fermi level, and the low- and middle-energy van Hove singularities. That is, such experimental examinations can provide the critical information on the multi-orbital hybridizations in the p–spScanning tunneling spectroscopy (STS) can provide sufficient information on the density of states at the Fermi level and the various van Hove singularities due to the valence and conduction bands simultaneously. High-resolution STS measurements are available for distinguishing the semiconducting and metallic behavior. Furthermore, they are very useful in identifying the close relations between the electronic energy spectra and the orbital hybridizations of the significant chemical bonds. Such experimental measurements have been successfully utilized to verify the band properties near the Fermi level and the dimension-diversified van Hove singularities in the graphene-related systems even in the presence of magnetic field, such as 2D few-layer graphene systems with the AB, ABC, AAB stackingsThe diverse structural and electronic properties of Si-doped graphene systems have been studied by the first-principles calculations. Apparently, the geometric structures, band structures, spatial charge densities and DOSs exhibit the rich and unique features, in which they are sensitive to the various adatom adsorption and substitution configurations. The critical multi-orbital hybridization mechanism and chemical bonding scheme are proposed to explain the Si-induced diverse physical and chemical phenomena. The calculated results clearly show that free carriers and energy gaps might be created by the Si-adsorptions and Si-substitutions, respectively. The Si-adsorbed graphene systems can remain in the planar graphene plane, in which the optimal position corresponds to the bridge site. The planar hexagonal structures clearly indicate a very small variation in the The Si-adsorptions and Si-substitutions thoroughly alter the unique band structure of pristine graphene systems, especially for the zero-gap semiconducting behavior and linear Dirac cone made by the 89. The many-body exchange and correlation energies, which come from the electron–electron Coulomb interactions, are calculated from the Perdew–Burke–Ernzerhof (PBE) functional under the generalized gradient approximation90. Furthermore, the projector-augmented wave (PAW) pseudopotentials can characterize the intrinsic electron–ion interactions. As to the complete set of plane waves, the kinetic energy cutoff is set to be 500 eV, being suitable for evaluating Bloch wave functions and electronic energy spectra. A vacuum space of 10 Å is inserted between periodic images to avoid any significant interaction. The first Brillouin zone is sampled by The geometric structures and electronic properties of Si-adsorbed and Si-substituted graphene systems are thoroughly explored using the density functional theory (DFT) implemented in Vienna ab initio simulation package (VASP)91, germanene92 and tinene systems93.Via delicate VASP calculations on certain physical quantities, the critical physical and chemical pictures, i.e., the multi- or single-orbital hybridizations in chemical bonds due to C-host and Si-guest atoms, can be achieved under a concise scheme. They will be useful in fully comprehending the fundamental physical properties. These important concepts are obtained from the adsorption- and substitution-diversified geometric structures, carbon- and silicon-dominated valence and conduction bands, the total charge distributions and their drastic changes after adatom adsorption or guest-atom doping and the atom- and orbital-decomposed density of states through detailed analyses. Also, such physical quantities could shed light on the significant differences between the atom adsorptions and substitutions, such as the metallic or semiconducting behaviors, the normal and irregular electronic energy spectra, and the complicated van Hove singularities, being attributed to the diverse chemical bondings. The developed theoretical framework can be conceivably generalized to emergent 2D materials, e.g., the chemical adsorption and substitution in layered silicene


By in situ electrical doping of a graphene template to suppress the charge transfer between C60 ad‐molecules and graphene, the layer‐by‐layer growth of a C60 film on graphene can be achieved. Under this condition, the graphene–C60 interface is free of Fermi‐level pinning; thus, barristors fabricated on the graphene–C60 interface show a nearly ideal Schottky–Mott limit with efficient modulation of the charge‐injection barrier. Moreover, the optimized C60 film exhibits a high field‐effect electron mobility of 2.5 cm2 V−1 s−1. These results provide an efficient route to engineering highly efficient optoelectronic graphene–OSC hybrid material applications.Controlling the growth behavior of organic semiconductors (OSCs) is essential because it determines their optoelectronic properties. In order to accomplish this, graphene templates with electronic‐state tunability are used to affect the growth of OSCs by controlling the van der Waals interaction between OSC ad‐molecules and graphene. However, in many graphene‐molecule systems, the charge transfer between an ad‐molecule and a graphene template causes another important interaction. This charge‐transfer‐induced interaction is never considered in the growth scheme of OSCs. Here, the effects of charge transfer on the formation of graphene–OSC heterostructures are investigated, using fullerene (C 60) thin films on graphene templates where charge transfer occurs is presented. The number of electrons transferred from graphene to C60 is controlled by in situ electrical gating of graphene during C60 deposition. When this electron transfer is suppressed, high‐crystalline C60 thin films are achieved for highly efficient optoelectronic applications.The growth behavior of fullerene (C With the introduction of these unique graphene–semiconductor interfaces, researchers have proposed various graphene–semiconductor hybrid optoelectronic devices such as field‐effect transistors (FETs), light‐emitting diodes, solar cells, photodetectors, and barristors.Graphene has excellent properties, so the possibility of integrating it with both inorganic and organic semiconductors has been intensively studied. Graphene–semiconductor heterostructures provide multifunctionality and desirable properties for scalable and flexible optoelectronic applications.Graphene is inert and is composed of a single‐atom‐thick layer, so it is a useful growth template for semiconductors, especially organic semiconductors (OSCs).60) thin film on a graphene template can be controlled by tuning charge transfer between them. The Fermi level (EF) of the graphene template determines the amount of charge transfer between the graphene and the C60 ad‐molecules, and this amount in turn affects the molecular dynamics of C60 on the graphene template. By finely tuning the EF of the graphene template, we induced layer‐by‐layer growth of highly ordered C60 films on graphene. Considering that the thin film's topological and crystalline features determine the optoelectronic properties of OSCs,60 films grown under optimized conditions exhibited a maximum field‐effect mobility of 2.5 cm2 V−1 s−1. Furthermore, a graphene–C60 Schottky junction prepared by our method approached the Schottky–Mott limit, which is desirable for highly efficient graphene–OSC barristors and other vertical graphene–OSC hybrid optoelectronic devices.Here, we demonstrate that an epitaxial growth of a vacuum‐deposited fullerene . To clarify the relationship between charge transfer and the initial electronic states of graphene, we fabricated graphene field‐effect transistors (G‐FETs) on 300 nm thick SiO2/Si substrates and compared the transfer characteristics of the G‐FETs before and after 3 s of C60 deposition at a deposition rate of 5 × 10−2 monolayer per second (ML s−1)  could be solely attributed to the change in charge‐carrier density .We first investigated the transfer of electrons from graphene to C1) Figure1a. To 60 deposition, the RCh was preserved as long as the gate voltage (VG) was <−40 V. This preservation demonstrates that deposition of C60 did not cause degradation of graphene, and more importantly, that no charge transfer occurred between graphene and C60 in this range of VG. However, at VG > −40 V, the RCh–VG curve shifted to the right; this change indicates that electrons were transferred from graphene to C60 .After C0 Figure b. This snCT (cm−2)) at a certain VG, the RCh–VG curve was fitted using the constant‐mobility model.60 deposition  and the carrier density of graphene–C60 after C60 deposition  were each calculated at each VG asVD is VG at maximum RCh, μ is the carrier mobility, e is the elementary charge, L is the channel length, W is the channel width, and nres is the residual carrier concentration in graphene. Then ΔnCT was calculated as ng,bare−ng,C60. Before C60 deposition, the fitted values of μ and nres of the graphene transistor were 4470 cm2 V−1 s−1 and 2.3 × 1012 cm−2, respectively. When plotted versus ng,bare .To calculate the number of transferred electrons  level of adjacent C60. The LUMO level of isolated C60 molecules is known to be −4.5 eV,EF of undoped graphene. However, the energy levels of organic molecules change and broaden upon adsorption of C60, because of the polarizability of the substrate;60 adsorbates can lie below the EF of undoped graphene that has ng,bare > nc; as a result, the graphene becomes p‐type doped. The absence of charge transfer when ng,bare < nc is attributed to the EF of graphene being lower than the LUMO level of the C60 adsorbates  that is shifted downward and an unoccupied LUMO level (L2) that is shifted upward  the growth behavior of C60 crystals as the EF of graphene gradually approached the EF,c.With the in situ electrical gating of graphene , we observed changes in i) the molecular interactions and assembly of C60 ad‐molecules may interact with each other on the graphene surface, depending on the relative position of the EF of graphene and the EF,c. These distinctions can be well detected by Raman spectroscopy . In both Raman spectra, the feature peaks of C60, i.e., the Ag(1) mode at ≈500 cm−1 and the Ag(2) mode at ≈1470 cm−1, were clearly observed.Ag(2) peak indicates the number of intermolecular bonds to each C60 molecule, where each intermolecular bond shifts the peak by −5 cm−1.Ag(2) mode of C60 grown on graphene with EF < EF,c is consistent with that reported for pristine C60 molecules.Ag(2) peak of C60 grown on graphene with EF > EF,c was red‐shifted ≈3 cm−1; this change indicates that chemically bonded C60 dimers or oligomers were formed. This selective formation at high EF strongly suggests that control of the EF of graphene during C60 growth indeed determined the charge state of the C60 ad‐molecules.First, C60 ad‐molecules determines the formation of covalent bonds between two C60 molecules.60 molecules have negative charges, the activation barrier for the bonding decreases. Therefore, graphene with EF > EF,c induced negative charges in C60 ad‐molecules, resulting in the formation of intermolecular bonds between C60 ad‐molecules. By contrast, on graphene with EF < EF,c, C60 molecules were charge‐neutral and thus did not form covalently bonded C60 dimers.The charge state of CnCT affected molecular arrangement in C60 crystals, and consequently, how those crystals assembled into thin films. We used grazing incidence X‐ray diffraction (GIXD) to characterize C60 thin films with different thicknesses grown on graphene, where ΔnCT was controlled. Under ambient conditions, the most stable structure of C60 crystals is face‐centered cubic (fcc);60 were observed in our system of C60 thin films grown on graphene  and at 30° and 35° tilt from zq, respectively. These results indicate that C60 has an epitaxial relationship with graphene, with the (111) plane of C60 crystals parallel to the graphene substrate;nCT. However, differences were observed in the crystal domain sizes of C60 thin films grown on graphene at different ΔnCT  (R(111)). When ΔnCT = 0 during C60 growth, C60 thin films had R(111) ≈ 60 nm, which is almost three times larger than in the film grown under very high ΔnCT.At the early growth stage , irrespective of the occurrence of charge transfer, the set of reflections of (111) family and the reflections of plane (113) and plane (220) appeared; these reflections are located along the out‐of‐plane direction  images  grown on graphene were imaged at high magnification using HR‐TEM. The image of C60 grown on graphene at ΔnCT = 0 clearly showed an ordered hexagonal arrangement of C60 molecules over a few tens of nanometers, which is the fashion of the (111) plane of a highly crystalline fcc structure  pattern of this C60 thin film also showed only a single set of hexagonal patterns, i.e., the crystalline orientation of C60 was uniform along the vertical direction. Notably, when ΔnCT = 0 was maintained during C60 growth, the misorientation angles between the SAED patterns of C60 and those of graphene were concentrated at close to 0° and 30°, which correspond to energetically stable adsorption sites of C60 molecules along the armchair and zigzag directions of graphene, respectively .60.Therefore, C60 was grown on graphene under a very high ΔnCT, HR‐TEM image and the corresponding SAED patterns  in this film  randomly oriented nucleation of C60 crystals and ii) the formation of additional amorphous layers, or both. The resulting richness of tilt grain boundaries could result in the observed polycrystallinity along both the lateral and vertical directions. The dominance of (111)‐plane‐oriented C60 crystal domains . To quantize the dimer content, we analyzed numerous intermolecular distances of two nearest‐neighbor C60 molecules in films grown at ΔnCT = 0 and ΔnCT > 0  levels.In addition, we performed density functional theory (DFT) simulations to calculate the electronic structure of a double‐bonded C60 growth behaviors are further demonstrated by morphological analysis using atomic force microscopy (AFM)  and a large increase of surface coverage. As electron transfer from the graphene to C60 ad‐molecules increased, the number of nuclei quickly increased and each of them merely grew in height; the result was an array of grains of different heights. At ΔnCT = 0, as the growth continued, continuous C60 film was formed by coalescence of large‐area C60 grains; by contrast, at ΔnCT > 0, C60 film was formed by full coverage of small C60 islands with poor inter‐grain connection. At the later growth stage (12.5 ML), the C60 thin film grown at ΔnCT = 0 revealed clear terrace structure, which is evidence of lateral growth mode, whereas the film grown at ΔnCT > 0 simply showed an array of tiny crystallites.The effects of charge transfer on CM) Figure4a, whi60 system as well as its effects on the crystal structure and morphology of C60  of these films against ΔnCT at room temperature revealed correlations between the nucleation of C60 and charge transfer from graphene to C60  as a function of ΔnCT, as Ni = C exp (ENuc/(kBT)) where C is a pre‐exponential factor, kB is the Boltzmann constant, and T is the substrate temperature.Ni values as a function of the substrate temperature T were collected at various fixed ΔnCT; the slopes of plots of ln(Ni) versus 1/(kBT) at a certain ΔnCT gave the values of ENuc at the ΔnCT .ENuc is related to the activation energies of all of these atomistic processes. However, the energy barrier for C60 diffusion is negligible on graphitic surfaces,60 on graphene are predominantly limited by the rate of attachment of ad‐molecules to pre‐existing islands.The nucleation of Ces Figure5a. AftENuc = [2Ei + 2(i + 1)EB]/(i + 3),i is critical cluster size, Ei is cluster energy, and EB is activation energy for the attachment.EB increases. This relation is explained as follows. The presence of high EB hinders the attachment of deposited ad‐molecules to an island, so the concentration of ad‐molecules increases on the graphene surface. Thus, the probability of ad‐molecules colliding rapidly increases, and this change favors new nucleation rather than the growth of pre‐existing islands.For such attachment‐limited nucleation with negligible barriers to diffusion and dimerization, Ni and ENuc with increasing ΔnCT are attributable to the increase in EB with increasing ΔnCT as EB (ΔnCT) = EB0  + E′B(ΔnCT) where EB0 is the charge‐transfer‐independent attachment barrier and E′B is the charge‐transfer‐dependent attachment barrier. When electrons in graphene are transferred to the ad‐molecules and the islands, the ad‐molecules and islands are negatively charged and the underlying graphene becomes positively charged  dominantly affects the nucleation kinetics. This assumption can be justified because the probability of collision between two C60 molecules which both simultaneously have negative charges would be very small. On the contrary, a C60 island contains many C60 molecules, so a C60 island is likely negatively charged.Therefore, the increases in 60 can be simply explained by invoking the repulsive Coulomb interaction between an ad‐molecule and an existing island. With increasing ΔnCT, EB increases because of the repulsive interaction; this change inhibits the lateral growth of negatively charged islands by negatively charged ad‐molecules diffusing on the graphene surface. However, irrespective of the EF of the graphene template, the ad‐molecules from the vapor phase can land directly on the top of the existing island because they are charge‐neutral and thus not prone to the repulsive Coulomb interaction. However, after they are deposited on the top of the islands, their dynamics are again influenced by the EF of graphene. When ΔnCT = 0, they can move relatively freely down to the graphene surface because the Ehrlich–Schwoebel barrier is much lower than the diffusion barrier on top of the C60 layer.60, the EB increases and thus acts as an energy wall surrounding the edge of islands. For an ad‐molecules on the top of the island to move downward and escape from the island, they must overcome an activation energy greater than EB. Consequently, ad‐molecules become concentrated on the top of the island, so the island rapidly grows in the vertical direction. The rapid vertical growth in turn leads to the formation of randomly oriented crystals.The transition from a 2D to a 3D growth mode Figures  under th2.460 thin films for lateral charge transport, we grew C60 thin films on graphene at controlled charge‐transfer conditions, then transferred the C60 films to octadecyltrichlorosilane (ODTS)‐treated SiO2/Si substrates and then fabricated planar C60 transistors (C60‐FETs). The final device included an ≈50 nm thick C60 channel without the underlying graphene  and measured the on/off ratio (Ion/Ioff). For a C60 thin film grown at ΔnCT = 0, the Ion/Ioff of the FET device was ≈107 and the average μe was ≈1.5 cm2 V−1 s−1. The maximum mobility of the device was ≈2.5 cm2 V−1 s−1, which is similar to the state‐of‐the art mobility of C60 transistors fabricated by the vapor deposition method  between the C60 layer and the bottom graphene. Increasing the VG barely affected the current in the forward regime (VDS < 0) but boosted the current in the reverse regime (VDS > 0). Consistent with the band diagram .Such charge‐injection efficiency at the graphene–C60 layer that had been grown at ΔnCT = 0 showed substantial modulation of the reversed current by the gate voltage; and the on‐state current Ion was higher in this device than in the device that used the C60 layer that had been grown at ΔnCT > 0, whereas their off‐state currents Ioff were similar. As a result, this device fabricated with a highly crystalline C60 film  achieved an Ion/Ioff ratio of ≈103 at VDS = 2 V, which is nearly two orders of magnitude greater than the Ion/Ioff ratio of the other device at the same VDS  required to induce charge transfer between graphene and C60  versus 1/(kBT)  large amorphous areas, and iii) other crystalline defects that would introduce numerous interfacial trap states . The Fermi level of graphene was pinned at those states. In addition, because the DFT results reveal a smaller bandgap of a C60 dimer compared with two isolated C60 molecules  were measured under light illumination at 0.62 eV . For comparison, C60 thin films were grown on top of graphene channels under ΔnCT = 0 and ΔnCT > 0. At a high positive gate bias (VG = 80 V), only the G‐FET with the C60 thin film grown at ΔnCT > 0 showed additional photocurrent as the device was illuminated . The excitation energy is much smaller than the bandgap of the C60 thin film and smaller than 2|EF| of graphene at VG = 80 V, so the interband transitions are forbidden in both the C60 thin film and the graphene.60 thin film was merely a result of detrapped electrons from the interfacial states, which were abundant in the layer grown at high ΔnCT. In fact, we observed positive photocurrent from the G‐FET with the C60 film grown at ΔnCT > 0, but observed no photoresponse from the device with C60 film grown at ΔnCT = 0.To directly confirm the interfacial states between the graphene and the C360 system during the growth of C60 crystals on a graphene template governed such growth and, thus governed the thin film's corresponding crystal structure and morphology. These charge‐transfer phenomena altered the electronic states of the graphene–C60 system, forming negatively charged C60 nuclei and ad‐molecules. Under these conditions, the growth of C60 on graphene was favored in the vertical dimension because of the high attachment barrier energy, resulting thin films with small and randomly oriented crystallites. With this understanding, we proposed that the optimized graphene template for layer‐by‐layer growth of C60 with large and uniformly oriented crystals is the graphene in which the charge transfer from graphene to C60 is suppressed during the C60 growth. Barristors fabricated with this graphene–C60 van der Waals heterostructure showed efficient tunability of the charge injection barrier, approaching the Schottky–Mott limit. In addition, the lateral electron mobility μe in a planar C60‐FET was also boosted to a maximum μe = 2.5 cm2 V−1 s−1.We observed that charge transfer within the graphene–CThe authors declare no conflict of interest.Supporting InformationClick here for additional data file.


The polarization insensitive modulator is designed to achieve a maximum experimental modulation depth of about 53% within 5 V of gate voltages using monolayer graphene. Moreover, the manufacturing processes are inexpensive. Two methods are adopted to improve modulation performance. For one thing, the metal metamaterial designed can effectively enhance the electromagnetic field near single-layer graphene and therefore greatly promote the graphene’s modulation ability in terahertz. For another, polyethylene oxide-based electrolytes (PEO:LiClO The electromagnetic spectrum from 0.1 to 10 terahertz (THz) is called the THz region. As they have superior intrinsic properties, terahertz waves can interact with various materials and have a wide range of applications, such as penetrating imaging , and med2V−1s−1) without a high incident excitation source at room temperature [Similar to the active modulators of optical and microwave regimes as reported, the established ones in THz are also in the demand for high efficiency, fast modulation speed, and low power consumption . Graphenperature . Therefoperature .MD) while lowering the drawing-voltage.Although some progress has been made on electrically tunable graphene modulators in THz, the performance of such modulators still needs enhancing. One of the basic focuses is on increasing the modulation depth (MD [The monatomic layer thickness of graphene (~0.36 nm) is disadvantageous for strong interactions with electromagnetic waves passing through, which will inevitably lead to a small MD ,11. TherMD ,13,14,15MD .−2) is ≈800 times higher than that of the 300 nm SiO2 plate capacitor [−2) [In addition, another deficiency is that the drawing-voltage of THz graphene modulators is generally as high as several tens of volts ,17. Giveapacitor ,19,20. Mtor [−2) ,24,25,26tor [−2) . Aside ftor [−2) ,29,30.MD of 53% is achieved within 5 V of gate voltages with this device. Besides, the fabrication of this device demands inexpensive equipment. This architecture provides a feasible approach for optimizing THz active modulation performance with single-layer graphene. Additionally, because the flower-shaped structure in this work is considered a kind of customized metamaterial, it could be replaced corresponding to a specific application in the future.In this work, we propose an effective THz configuration comprised of metamaterials, solid electrolyte, and graphene to realize dynamical modulation of device transmittance. By using the solid electrolyte top gate, we can obtain a highly doped graphene membrane with activity tunable carrier densities via small gate voltage. Moreover, it has been experimentally verified that a maximum The schematic of the THz amplitude modulator is shown in 4), are employed to cover the modulation region and gate electrodes. Voltages applied on the top gate can electrolyze solid electrolyte into Li+ and ClO4− ions [12 cm−2) [fE) in the monolayer graphene, can be significantly changed using drawing-voltages [The central functional section of this modulator is composed of five layers from the bottom to the top, as demonstrated in O4− ions , and the12 cm−2)  and Fermvoltages .Structures of the fabricated functional region are shown in 2O, acetic acid, and nitric acid , with a volume ratio of 16:2:1:1. By carefully selecting the immersion time, the depth and width of etched trenches could be controlled to form accurate and uniform large-area structures, as presented in Firstly, the metallic structure and electrodes are fabricated through wet etching. Specifically, PET with 500 nm thick aluminum, the raw material bought directly , was spin-coated with 1 μm thick S1805 photoresist  and exposed to ultraviolet light under a mask , to pattern the photoresist with flower-shaped structures. Then, the membrane with developed photoresist was immersed into the corrosive liquid  for 80 s to remove the unwanted part of the aluminum layer. The etchant mentioned was comprised of phosphoric acid, HSecondly, an alumina spacer  with a thickness of 50 nm was deposited on metamaterial film by electron beam evaporation . In the practical process, a hard mask was used to shield the electrodes.2/Si. One can see that the D peak (≈1300 cm−1) indicates defects are extremely low, and the intensity ratio of G (≈1600 cm−1) to 2D peak (≈2700 cm−1) is 1:2, which satisfies the characteristics of SLG [Thirdly, a commercially chemical vapor deposition (CVD)-grown graphene strip  was carefully transferred via the poly(methyl methacrylate) -assisted wet-transfer process onto the dielectric layer to connect the source with drain electrodes . Given ts of SLG ,36. The TGV) varying from −10 V to 6 V, while the drain voltage was fixed at 0.1 V. Therefore, the FET function of the device was confirmed to be effective. The charge neutrality point (CNP) shifts to a negative value, because the graphene flake with higher work function acts as an acceptor of electrons (n-dopants) when it contacts electrodes. According to the report [Finally, the ion-gel of 100 nm thickness, whose composition is referred to in the literature  , is spine report , the worMD is a crucial indicator, which is defined as:VTg is the measured transmittance of a sample at a specific gate voltage, and maxT is the maximum transmittance among different voltages. The experimental MD is demonstrated in maxT occurring at −3.5 V, the maximum MD is 55% at 6 V. The relationship between gate voltage and MD is non-linear. In detail, the increased rate of MD gradually slows down, with the applied voltages shifting away from the Dirac point.The modulation capability of this device was investigated with a THz-TDS system, from 0.2 to 1 THz . The orifE is the chemical potential of the graphene, e = 1.602 × 10−19 C is the electron charge, BK is the Boltzmann’s constant, T = 300 K is the temperature, and To analyze the device theoretically, we utilize the finite element method to establish a theoretical model of the device. Due to the periodic metamaterial in the functional region, the original model is simplified and represented by one cell with a periodic boundary in the horizontal plane. As for structure sizes, they all refer to the actual ones of the modulator, as shown in lumina=6 , and thelyte=5.5 . The con formula :(2)σgrapIn order to analyze the device using a theoretical model, we need to figure out the relationship between the applied gate voltage and the corresponding Fermi level of SLG graphene. The electrolyte capacitance is the key cue to solving this problem.4 [gC and SiOC2 are the capacitance of the solid electrolyte and the 300 nm SiO2/Si (0.0121 μFcm−2) respectively [BG,CNPV is the back-gate voltage of CNP. TGV, the Dirac point moves to left gradually. The specific movement rule of CNP is shown in 2 plane-parallel capacitor. Eventually, the free carrier concentration of electrolytes under various TGV could be theoretically predicted by the following equation [Vg is gate voltage, CNPV is the CNP voltage, 6 m/s [n is the actual carrier density. Moreover, when Equation (3) combines with Equation (4) below, the fE could be calculated as a function of the gate voltage in We adopted a so-called dual-grating-gate configuration, demonstrated in 4 . As the ectively ,19,20, aequation ,38.(3)V 106 m/s , and n i2/Si, solid electrolyte requires lower voltage at the same doping level. Particularly, the TGV of −10 V, −4.5 V, 0 V, and 5 V corresponds to the chemical potential of 0.39 eV, 0.15 eV, 0.28 eV, and 0.45 eV, respectively.According to In this section, we aim to prove that the proposed metallic structure can indeed promote modulator performance with single-layer graphene.MD of the device with the metamaterial layer was deeper than the one without at resonance frequency. To be specific, in 0.48 THz, the MD rises from 11.6% to 75.4% within 0.4 eV after employing the metamaterial layer.In fE of graphene is 0 eV, and the frequency is 0.48 THz in the parameter settings. It was observed that the local field intensity near the metallic slits was enhanced, with an average of 102 orders, compared with that of the control group. Further study showed that, accompanied by the gradual increasement in the Fermi level, the resonance coupling strength receded evidently.Moreover, the enhancement ratio of the electric field in the graphene layer was demonstrated in In conclusion, the proposed metamaterial can effectively strengthen the local electric intensity around the SLG at the resonance frequency. Consequently, the dynamic modulation ability of SLG can be magnified by the enhanced field. Therefore, the flower-shaped structure benefits the deep modulation of THz transmittances by monolayer graphene.fE in fE of graphene, the theoretical transmittance is slightly higher than the measured ones, because of the non-loss material settings of the solid electrolyte and the dielectric layer in the analytical model. According to Experimental transmission spectra are compared with simulated ones with a higher level of Additionally, the summits of transmittance versus voltages are illustrated in T) and electric resistance of a graphene flake (R) could be described by Equations (5) and (6) for a FET-type modulator [Cg is the gate capacitance of FET, μ is the carrier mobility, Vg is the gate voltage, DiracV and DiracR are the corresponding voltage and resistance of graphene in Dirac point, and L and W are the length and width of the graphene strip, respectively.According to previous studies, the relationship between transmittance (odulator ,45,46,47DiracV, which is in good agreement with Vg applied shifts away from the DiracV, the transmittance would gradually decrease.Combining the two equations above, we could conclude that the transmittance reaches the maximum at a gate voltage equal to The peak values of theoretical transmittances are extracted and drawn in black lines, while the experimental ones are shown in black dots. It was noted that the lines were only consistent with dots when the graphene Fermi level was higher than 0.17 eV. In other words, the experimental CNP no longer corresponds to 0 eV, and the Dirac point is no longer a single dot. As a consequence, the maximum transmittance of the experiment is only about 30% in In summary, we propose a top-gated THz device to realize the high-performance modulation of single-layer graphene. In order to improve the poor performance induced by weak interaction and large applied voltages, this modulator employs a metallic metamaterial and solid electrolyte layer to target the flaws. Specifically, the enhanced electric field excited by the flower-shaped structure can strengthen the interaction when the terahertz wave goes through the single-layer graphene, and the solid electrolyte can effectively elevate the doping level of graphene at the same gate voltage. Due to these two optimizations, the modulator efficiency was notably increased. It was verified that the maximum modulation depth is ≈78.7% in theory and ≈53% in experiment within 5 V of gate voltages in this paper. Besides, the fabrication of the device is also inexpensive. The metamaterial layer in this work could be replaced by a customized one for a specific application in the future.


V1.3 and KCa3.1 for SMC proliferation in arteriogenesis. Employing a murine hindlimb model of arteriogenesis, we found that blocking KV1.3 with PAP-1 or KCa3.1. with TRAM-34, both interfered with reperfusion recovery after femoral artery ligation as shown by Laser-Doppler Imaging. However, only treatment with PAP-1 resulted in a reduced SMC proliferation. qRT-PCR results revealed an impaired downregulation of α smooth muscle-actin (αSM-actin) and a repressed expression of fibroblast growth factor receptor 1 (Fgfr1) and platelet derived growth factor receptor b (Pdgfrb) in growing collaterals in vivo and in primary murine arterial SMCs in vitro under KV1.3. blockade, but not when KCa3.1 was blocked. Moreover, treatment with PAP-1 impaired the mRNA expression of the cell cycle regulator early growth response-1 (Egr1) in vivo and in vitro. Together, these data indicate that KV1.3 but not KCa3.1 contributes to SMC proliferation in arteriogenesis. Collateral artery growth (arteriogenesis) involves the proliferation of vascular endothelial cells (ECs) and smooth muscle cells (SMCs). Whereas the proliferation of ECs is directly related to shear stress, the driving force for arteriogenesis, little is known about the mechanisms of SMC proliferation. Here we investigated the functional relevance of the potassium channels K Arteriogenesis, which is defined as the growth of pre-existing arteriolar connections into functional arteries compensating for the loss of an artery due to occlusion , particuEgr1) [SMCs, located at the abluminal side, are not able to sense shear stress, and relatively little is known about the mechanisms triggering SMC proliferation in arteriogenesis. Growth factors, such as fibroblast growth factor 2 (FGF-2) and platelet-derived growth factor BB (PDGF-BB) have been shown to be important for SMC proliferation in arteriogenesis ,7 (and oEgr1) , a transEgr1) . InteresEgr1) . These dEgr1) ,12. Egr1. [Egr1, which is tightly regulated by the transcriptional co-activators myocardin and myocardin-related transcription factors (MRTFs) and the ternary complex factor ETS like protein Elk-1, which compete for the binding to the transcription factor serum response factor (SRF) [The proliferation of vascular SMCs in arteriogenesis is characterized by the transition from the contractile to the synthetic (proliferating) SMC type. This process is associated with reduced mRNA and protein levels of contractile genes such as α-smooth muscle actin (αSM-actin) and paralleled by an increased expression of Egr1. ,13,14. Tor (SRF) ,16,17, hor (SRF) . V1.3 and KCa3.1 have been shown to be involved in cell cycle regulation by activating intracellular signaling pathways [V1.3 and KCa3.1 to the proliferation of vascular SMCs in collateral artery growth. The mode of action of these potassium channels in terms of proliferation is still under debate and several mechanisms, either ion flux dependent or independent, have been proposed [V1.3 and KCa3.1, have been demonstrated to be upregulated in proliferating SMCs, whereby their specific blockade interfered with cell cycle progression. Increased expression levels of KV1.3 have been detected in vitro in proliferating SMCs isolated from murine femoral arteries or from human donors [V1.3 with selective blockers such as 5-(4-phenoxybutoxy)psoralen (PAP-1) inhibited migration and proliferation of SMCs in vitro [V1.3, KCa3.1 has been found to be upregulated upon stimulation with PDGF-BB in proliferating SMCs in vitro [Ca3.1 with the selective blocker TRAM-34 in contrast interfered with SMC proliferation both, in vitro and in vivo [Ca3.1 plays a major role in endothelium-derived hyperpolarizing factor (EDHF)-mediated vasodilation as shown in KCa3.1 deficient mice [The potassium channels Kpathways ,19, and pathways ,21. Sincproposed ,19. Bothn donors  as well n donors ,23. Morein vitro ,23 and iin vitro . Similarin vitro  and in min vitro ,26. Bloc in vivo ,25,27,28ent mice . V1.3 and KCa3.1 for SMC proliferation in growing collateral arteries by performing blocking studies employing the selective channel blockers PAP-1 and TRAM-34, respectively. From our results we conclude that KV1.3 contributes to SMC proliferation in arteriogenesis, whereas KCa3.1 is more likely to be involved in vasodilation.In the present study we investigated the relevance of the potassium channels KMale C57BL6/J mice, purchased from Charles River, were housed in cages and kept under 12 h day/night cycle with food and water ad libitum. All experiments were approved by the Bavarian Animal Care and Use Committee  and carried out according to the guidelines of the German law for protection of animal life. Mice at the age of 6 to 10 weeks were anesthetized with a combination of 0.5 mg/kg medetomidine (Pfister Pharma), 5 mg/kg midazolam (Ratiopharm GmbH), and 0.05 mg/kg fentanyl (CuraMED Pharma). Arteriogenesis was induced by right femoral artery ligation ), whereas the left femoral artery was sham operated  as previV1.3 channel blocker (5-(4-phenoxybutoxy)psoralen , Sigma-Aldrich) [Ca3.1 channel blocker TRAM-34  [To block potassium channels, mice were treated either with the selective KAldrich) , or the ne Labs) , dissolvne Labs) ,31. The The laser Doppler perfusion measurements were performed as described in . In briePrior to tissue sampling for (immuno-) histology, mice were perfused with an adenosine buffer , both from Sigma-Aldrich dissolved in PBS, PAN Biotech, pH 7.4) for maximal vasodilation followed by perfusion with 3% paraformaldehyde  dissolved in PBS, pH 7.4, for cryoconservation, or 4% PFA for paraffin embedding . For qRTMouse primary artery smooth muscle cells  were cultured in a SMC growth medium  containing insulin and the growth factors fibroblast growth factor 2 (FGF-2) and epidermal growth factor (EGF) together with 20% fetal calf serum . For serum starvation, cells were cultured in Dulbecco’s modified Eagle´s medium  with 1% FCS for 24 h. Thereafter, negative controls were stimulated with medium containing 2% FCS, positive controls with 10% FCS.Giemsa staining on paraffin fixed tissue samples was performed according to standard procedures, and slices were analyzed using an Axioskop 40 microscope (Carl Zeiss AG). BrdU staining of paraffin fixed tissue sections was performed with a BrdU detection kit (BD Pharmingen) according to the manufacturer´s procedure using the same microscope for evaluating the tissue sections.To investigate the proliferation of mouse primary artery SMCs, a BrdU proliferation assay kit (Roche) was used according to the manufacturer´s instructions. In brief, mouse primary artery SMCs were seeded in a 96-well plate overnight, and after serum starvation in DMEM containing 1% FCS for 24 h, the mouse primary artery SMCs were cultured in DMEM with 10% FCS and treated with or without PAP-1 or TRAM-34, respectively, together with 10 mM BrdU. Cell proliferation was assessed by colorimetry with an Infinite F200 ELISA reader (TECAN).V1.3  or a rabbit anti-KCa3.1  antibody (both from Alomone Labs) followed by a goat anti-rabbit IgG Alexa fluor 488-conjugated antibody  together with a Cy3-conjugated mouse anti-αSM-actin antibody  and an Alexa fluor 647-conjugated rat anti-CD31 antibody , followed by DAPI counter staining . Images were taken with an Axio Imager 2 fluorescence microscope equipped with and an Axion ICc 5 camera and Axiovert software (Carl Zeiss) or using a LSM 880 confocal laser scanning microscope equipped with an Airycan module (Carl Zeiss) with ZEN black software for imaging acquisition. Imaging analysis of KV1.3 or KCa3.1 expression in αSM-actin positive SMCs or CD31 positive ECs was performed using the ZEN blue software. For the colocalization anaylsis, the ZEN colocalization tool was used (Carl Zeiss AG). The three-dimensional (3D) projection surface reconstruction of the images where done by using the Imaris software (Bitplane).For immunofluorescence staining, cryofixed tissue sections (10 μm) were stained with a rabbit anti-K® Reverse Transcription Kit (Qiagen) according to the manufacturer´s procedure. The qRT-PCR was performed as previously described [Fgfr1) forward 5′-CTTGCCGTATGTCCAGATCC-3′, reverse 5′-TCCGTAGATGAAGCACCTCC-3′, platelet derived growth factor b (Pdgfrb) forward 5′-AGGACAACCGTACCTTGGGTGACT-3′, reverse 5′-CAGTTCTGACACGTACCGGGTCTC-3′, early growth response 1 (Egr1) forward 5′-CGAACAACCCTATGAGCACCTG-3′, and reverse 5′-CAGAGGAAGACGATGAAGCAGC-3′. To control specific amplification, melt curve analyses and agarose gels were performed. Data were analyzed using the ΔΔCt method [The total RNA was isolated from the mouse primary artery SMCs or collateral arteries with Trizol (Life Technologies) and the residual DNA was removed by digestion with RQ1 RNase-Free DNase (Promega). Thereafter, RNA was purified with RNeasy MinElute columns (Qiagen) and reverse transcribed to cDNA using the QuantiTectescribed  using tht method  and resup ≤ 0.05.Statistical analyses were performed using the GraphPad software PRISM6. All data are stated as means ± SEM. Results were tested for normality and statistical analyses were performed as specified in the figure legends. Results were considered to be statistically significant at V1.3 or KCa3.1, respectively, were expressed in adductor collateral arteries. Immunofluorescence imaging revealed that KV1.3 and KCa3.1 labelling strongly colocalized with αSM-actin, a marker for SMCs, but weakly with CD 31, which is a marker for ECs  or TRAM-34 , respectively. Interestingly, in in vitro, both PAP-1 and TRAM-34 treatments interfered with SMC proliferation, as shown by the BrdU incorporation assay  revealed several binding sites for the transcription factor specificity protein 1 (Sp1) in the promoter regions of Fgfr1, Pdgfrb, and Egr1. However, whether Sp1 is indeed involved in potassium channel Kv1.3 mediated gene expression remains to be determined by further studies. Our data indicate that Kv1.3 plays a major role in SMC proliferation, especially in the process of arteriogenesis, by influencing signal transduction cascades associated with the expression of the growth factor receptors Fgfr1 and Pdgfrb and their downstream genes being involved in phenotype switch and cell cycle regulation.Previous results have shown that SMC proliferation in arteriogenesis is dependent on the activation of FGFR-1. Already in 2003, we had demonstrated that FGFR-1, which was expressed in SMCs but not in ECs, was upregulated in the early phase of arteriogenesis, i.e., within the first 24 h after induction of collateral artery growth by femoral artery ligation, and that blocking this tyrosine kinase receptor with polyanetholsulfonic acid (PAS) interfered with the process of arteriogenesis . A paraland Egr1 , whereasand Egr1 . AccordiCa3.1 selective blocker TRAM-34 did not influence vascular SMC proliferation or differential gene expression in growing collaterals in vivo  and in SMC proliferation, e.g., during atherogenesis [Ca3.1 is not involved in EC or in SMC proliferation in collateral artery growth  has previously been shown to play a role in arteriogenesis by promoting vascular cell proliferation [2+ levels [2+ levels could result in EDHF-mediated vasodilation, whilst a prolonged raise could activate transcription factors causing vascular cell proliferation [Ca3.1 is involved in this EDHF mediated vasodilation, but further studies are necessary to investigate this assumption.In contrast to our findings regarding PAP-1 administration, treatment of mice with the K in vivo . The las in vivo . Interesr stress , the drir stress ,3. Moreoogenesis . Our stuy growth . Of courerplasia  and atheerplasia . Togetheigations . As our igations , which wigations , we hypo channel ,40. Howe channel . In termferation . TRPV4 i+ levels . However+ levels ,44,45. Iferation ,45. It iv1.3, but not KCa3.1, contributes to SMC proliferation in arteriogenesis by controlling the expression of growth factor receptors, as well as their downstream genes relevant for phenotype switch and cell cycle progression.From our investigations, we conclude that the potassium channel K