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. 2010 Dec 28;31(5):983–997. doi: 10.1128/MCB.00137-10

FIG. 2.

FIG. 2.

H-Ras forms a complex with Cdc42 on endosomes. (A) A representative clone of HTC75 cells stably coexpressing Yc-Cdc42 and either Yn-EM-H-Ras(12V) (green) or Yn-EM-H-RasΔE(12V) (white) was analyzed by FACS (top). The cells were lysed, and the protein levels were examined by Western blotting using an anti-GFP (α-GFP) antibody, which cross-reacts with YFP (bottom). (B) The same cells as in panel A were transfected with vectors expressing either CFP-tagged Rab5A or GalT, which mark endosomes and the Golgi compartment, respectively, and examined by confocal microscopy. YFP and CFP signals were pseudocolored green and red, and the outline of the cell was marked by a dotted line in the merged image. Scale bar, 10 μm. (C) Two clones each of HTC75 cells stably coexpressing Yc-Cdc42 and either Yn-H-Ras(12V) (green) or Yn-H-RasΔE(12V) (white) were analyzed by FACS, and protein expression levels were examined by Western blotting as described for panel A. (D) Cells in panel C were transfected with vectors expressing organelle markers and examined as described for panel B.