Cell fractionation SAS cells were seeded into 10cm dishes at 2x 106 cells/dish and cultured in DMEM for 18–24 h. The cells were starved for 24 h in serum-free media. After treatment with HGF for a given time, the cells were harvested and washed with ice-cold PBS. The cells were then resuspended in 100 lL of lysis buffer (10 mM Hepes, 10 mM NaCl, 0.1 mM EDTA, 0.1 mM EGTA, 1% NP-40, 0.5 mM phenylmethylsulfonyl uoride, 0.1 mM dithiothreitol, 0.1 mM sodium orthovanadate, and protease inhibitors) and incubated on ice for 10 min. The nuclei were collected by centrifugation at 2000 g for 5 min at 4 o C. The supernatant was collected as a cytosolic fraction. Protein concentration of each sample was determined.