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. 2008 Mar 20;27(8):1183–1196. doi: 10.1038/emboj.2008.54

Figure 2.

Figure 2

RME-2 is lost from cortical endosomes in rme-4 and rme-5 mutants. (A–A″, B–B″, C–C″, D–D″, E–E″, F–F″) Dissected gonads were immunostained with anti-RME-2 antibody. (A–A″) wild-type; (B–B″) rab-11(RNAi); (C–C″) rme-4(b1001); (D–D″) rme-5(b1013); (E–E″) rme-4(b1001) rab-11(RNAi); (F–F″) rme-5(b1013) rab-11(RNAi). All strains expressed YP170–GFP. Middle focal planes of oocytes are shown. Arrowheads in (E′) and (F′) indicate tubular structures aggregated in small patches of the oocyte cortex (see text). Bar, 10 μm. Fluorescence intensity of RME-2 staining as a function of position was graphed along a line through the centre of the oocytes as indicated in the paired panels (A″–F″). Arrows indicate the relative position of the oocyte plasma membrane. At least 10 animals for each strain were analysed and representatives are shown. (G, H) Immunoelectron microscopy of RME-2 was performed in wild-type N2 (G) and rme-5(b1013) mutants (H). In the cortical area of wild-type oocytes, RME-2 localizes to the plasma membrane and tubular and vesicular structures (endosomes). In rme-5 mutants, the RME-2 signal, and the number of yolk-containing vesicles were significantly reduced. Bars, 200 nm. (I) Quantitative analysis of RME-2 localization in oocytes. The number of gold particles in the cortical or central regions of oocytes per unit area were counted, and the ratio as compared with wild-type controls is shown (see Materials and methods). (J) RME-2 protein level is not reduced in rme-4 and rme-5 mutant animals. Total lysates were prepared from the strains indicated and were probed with anti-RME-2 antibody in western blots.