Cytohesin-1 phosphoinositide binding is required for membrane ruffling and HGF-dependent cell migration. (A) Confocal images of cells counterstained with phalloidin (F-actin) and DAPI, with or without HGF. (B) Kymographs were generated from linescans of the cells’ leading edge imaged between 15 and 60 min after HGF treatment. (C) Quantification of experiments shown in A. (D and E) Quantification of experiments shown in B. (F) Random cell migration of diglycine EGFP-CYTH1 or R280C-ovexpressing cells treated with or without HGF. (G) Confocal images of cells counterstained with phalloidin (F-actin) and DAPI and treated with or without HGF (H) Kymographs were generated from linescans of the cells’ leading edge imaged between 15 and 60 min after HGF treatment. (I) Quantification of experiments shown in G. (J and K) Quantification of experiments shown in H. Scale bars, 20 µm. Arrowheads indicate peripheral membrane ruffles. All quantified data indicate mean values ± SEM from three independent experiments. **, P < 0.01; ***, P < 0.001; n.s., not significant.