Figure 4

Effect of anti-HGF neutralizing antibody on c-Met signaling in PC-3 cells A, PC-3 cells were serum-starved overnight and treated with the anti-HGF neutralizing antibody (anti-HGF) or control antibody (nIgG) for 2 h before stimulation with or without HGF (25 ng/ml) for 15 min. B, PC-3 cells were serum-starved over night and continuously treated with anti-HGF or nIgG for 24 h. C, PC-3 cells were serum-starved overnight and rinsed twice in a wash-out buffer (serum-free medium with 0.5 M NaCl, pH 3.7) for 30 s. Whole cell lysates were harvested immediately followed by Western blot analysis. Actin (A, B) or total c-Met (C) was used as a loading control.