Fig. 4.

Inhibition of Met phosphorylation, cell proliferation, and migration by HGF mutants. (A) HGF mutants (0.003–25 μg/ml) together with HGF (50 ng/ml) were added to A549 cell layer for 10 min at 37°C. Met phosphorylation by HGF mutants V495G (▿), D672N (▵), and G498I (♦) was quantified by using a KIRA assay and normalized to WT HGF activity (control). (B) Proliferation of BxPC3 cells was measured with 0.025 μg/ml HGF together with 2 μg/ml (gray bars) and 20 μg/ml (black bars) of HGF mutants. (C) MDA-MB435 cell migration was carried out in transwell plates in the presence of 0.1 μg/ml HGF together with HGF mutants at 20 μg/ml. HGF mutants were compared with WT HGF by using Student's t test: *, P < 0.05; **, P < 0.005.