HGF recruits c-Met, ITGB4, and S1PR1 to lipid rafts.
A, Optiprep fractions were isolated and subjected to immunoblotting with an anti-cav-1 antibody. The 30% Optiprep fraction represents the lipid rafts. B, HPAEC were treated with HGF (25 ng/ml) for 1 and 5 min and the 30% Optiprep fraction was isolated and subjected to immunoblotting with antibodies specific for c-Met, ITGB4, or S1PR1. Phosphorylation corresponding to the same molecular weight was determined via repeat immunoblotting on the same membrane with specific antibodies as shown (representative blots shown, densitometry data expressed as fold change phosphorylation relative to control and normalized to total protein, n = 3/condition, *, p < 0.05 compared with untreated controls).