Fig. 1. Localization of Rab5 and its functional inhibition with Rab5N142I. (A) EGFP fluorescence was observed in longitudinal section of the eye from Rh1-GAL4/UAS-EGFP Rab5 fly. R, retina; L, lamina. (B-D) Immunoelectron microscopy using an anti-GFP antibody identified subcellular localization of EGFP-Rab5 in the cell body (B,C) and the nerve terminal (D) of photoreceptor cells. Scale bars, 100 µm (A), 200 nm (B,C), 400 nm (D). (E) Antigen specificity of anti-Rab5 antiserum. (Top) CBB staining of Drosophila Rab2, Rab3, Rab4, Rab5, Rab6 expressed in E. coli. (Bottom) Immunoblot analysis using the anti-Rab5 antiserum against the above Rab proteins. (F) Heads homogenate was fractionated by velocity sedimentation. Fractions containing synaptotagmin (Syt), syntaxin (Syx) or Rab5 were identified by immunoblotting. (G) The numbers of MVBs and Golgi bodies were counted on electron micrographs of photoreceptor cells from wild-type (open bars) or hs-GAL4/UASRab5N142I mutant (shaded bars) flies.

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