Fig. 3. Localization of ectopic H-Ras-GTP in non-starved CHO cells. CHO cells were transfected with GFP-RBD (51-131) in (A) or cotransfected with H-Ras (B-D) and fixed 24 h later. (A) The GFP-RBD is diffusely localized throughout the cytoplasm and is not clearly associated with any intracellular structure or feature at the plasma membrane. Although the GFP-RBD can be occasionally seen in small membrane ruffles, this is probably due to the nonspecific trapping of fluorescent protein since GFP alone can produce a similar localization. Single confocal z-section shown. (B) Co-transfection with H-Ras causes the recruitment of the GFP-RBD to perinuclear structures, and plasma membrane ruffles, in non-starved cells. Single confocal z-section shown. (C) Immunostaining for total Ras expression. (D) Maximum projection overlay of eight equally spaced z-sections from the dorsal to the ventral cell surface showing co-localization of GFP-RBD (green) and H-Ras (red).