Fig. 1. R-Ras uses endomembranes to reach the plasma membrane. Hela (A-C) and Cos-7 (D-F) cells were transfected with vectors pEGFP-R-Raswt (A,B) and pEGFP-R-Ras43N (C,D), or double transfected with pEGFP-R-Ras43N and pcDNA4/TO-Arf6-27N (E,F). EGFP-R-Raswt colocalized with a p115 Golgi marker (arrow) and was also seen on the nuclear envelope (arrowhead). In addition to these organelles, EGFP-R-Ras43N was found on vesicular (C) and vacuolar (D) structures, which colocalized with Arf6-27N (arrowheads) (E,F). HeLa cells were transfected with pEGFP-RRaswt (wt), pEGFP-R-Ras38V (38V), pEGFP-R-Ras43N (43N) and pEGFP-C1A, and total cell extracts were analysed by western blot (G). The positions of molecular size markers are indicated.