FIGURE 7.

HGF stimulates EGFR phosphorylation, but EGFR signaling does not affect HGF-dependent phosphorylation of c-Met. A, Suit-2 cells were plated on laminin-1-coated plates for 3 h and then treated with EGFR TKI (PD153035) for 30 min prior to 5 or 30 min of stimulation with 50 ng/ml HGF where noted. EGFR or c-Met were immunoprecipitated from cell lysates and immunoblotted for phosphotyrosine (PY20), EGFR, or c-Met as indicated. B, Suit-2 cells were plated on laminin-1-coated dishes as in A and stimulated with 5 ng/ml EGF or 50 ng/ml HGF (HGF50) or 100 ng/ml HGF (HGF100) for 5 min as indicated before harvesting immunoblot analysis. C, Panc-2G6, Panc-3D7, AsPC-1, and Suit2 cells were seeded into 60-mm dishes, serum-starved overnight, and then stimulated with 50 ng/ml HGF for 5 min. Cell lysates were then immunoblotted with EGFR phosphotyrosine antibodies as noted (B and C). Total EGFR and actin were used as controls.