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Fig. 6. Ectopic lateral PAR-2 in cul-2(RNAi) embryos. (A) The initial localization of lateral PAR-2, as monitored in epifluorescence movies of cul-2(RNAi) embryos expressing PAR-2::GFP and ß-tubulin::GFP (left), and PAR-2::GFP and histone H2B::GFP (right). Green arrows denote cortical PAR-2. Cortical PAR-2::GFP signals were observed in regions that were distal to the meiotic spindle (left), or to oocyte and sperm DNA (right), during the entire time period post-fertilization. (B) Epifluorescence (top) and confocal three-dimensional reconstruction of a cul-2(RNAi) embryo showing a circular PAR-2 pattern: PAR-2::GFP (green), anti-{alpha}-tubulin (red) and DAPI (blue). Note that the confocal reconstruction is only of the embryo and excludes the surrounding germ cell DAPI signal. Middle image shows a side view; bottom image, anterior is to the front. (C) Simultaneous localization of PAR-2 and PAR-6 in meiosis II-stage cul-2(RNAi) embryos. (Left) Anterior and lateral PAR-2::GFP (top) with uniform anti-PAR-6 staining (bottom). (Right) Anterior anti-PAR-2 staining with anterior exclusion of PAR-6::GFP. (D) PAR-2::GFP localization in tbb-2(RNAi) and tbb-2; cul-2 double RNAi meiotic embryos. Epifluorescence images of PAR-2::GFP (green, top), DAPI (blue, top) and anti-{alpha}-tubulin (red, bottom). o, location of maternal DNA; s, location of sperm DNA. The left tbb-2; cul-2 double RNAi embryo has a lateral PAR-2::GFP patch, whereas the right embryo has PAR-2::GFP on the end of the embryo near both the maternal and condensed sperm DNA. (E) emb-30(tn377) (left) and emb-30(tn377); cul-2(RNAi) (right) zygotes were stained with anti-{alpha}-tubulin (red) and either anti-PAR-2 (green, upper panel) or anti-PAR-6 (green, lower panel). For each genotype, left panels show anterior PAR-2 localization (top) or PAR-6 exclusion (bottom), and right panels show lateral/posterior PAR-2 localization (top) or PAR-6 exclusion (bottom). White arrows denote regions of the cortex lacking PAR-6 staining. Anterior is to the left. Scale bars: 10 µm.





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