Fig. 2. Plasma membrane localization of R-Ras molecules. HeLa cells transiently expressing EGFP-R-Raswt (A), EGFP-R-Ras38V (B), EGFP-R-Ras43N (C) and EGFP (D) were fixed and analysed by fluorescence microscopy. Note that EGFP-R-Raswt and EGFP-RRas38V are found in adhesion-like structures, whereas EGFP-RRas43N and EGFP are not. Localization of endogenous R-Ras (E) to vinculin (F) containing focal adhesions in untransfected HeLa were seen using specific antibodies against R-Ras and vinculin. EGFP-RRas38V was also detected in focal adhesions in C2C12 cells (G,H). Arrowheads indicate focal adhesion-like localization. Bars, 10 µm. (I) The percentage of cells containing R-Ras in focal adhesions was calculated for R-Raswt, R-Ras38V and R-Ras43N. Values are means ± s.d. from three independent experiments in which >100 cells were scored per condition.