Figure 7
Combined RNAi with the chitin-binding proteins, CEJ-1 and B0280.5, results in a gna-2(qa705) -like Pod phenotype. (A) CEJ-1 and B0280.5 have multiple chitin binding Peritrophin-A domains and mucin-like regions. Amino terminus is on the left. (B) Hatchling number of wild type, cej-1(RNAi), B0280.5(RNAi) or cej-1+B0280.5(coRNAi) (bars are mean ± SEM, n = 5, x,y: significantly different, p < 0.05). (C) DNA (Hoechst) and chitin staining of wild type or cej-1+B0280.5(coRNAi). (D) Wild type or B0280.5(RNAi) (controls), or cej-1+B0280.5(coRNAi) stained for DNA (Hoechst; blue) and PAR-3 (red), PGL-1 (pink) or PIE-1 (green). Images are overlays of Hoechst and antibody staining. (E) Osmotic sensitivity of cej-1+B0280.5(coRNAi) embryos expressing histone H2B::GFP, dissected in water, 150 mM KCl or 300 mM KCl. Images are overlays of histone H2B::GFP (chromatin; green) and DIC. (F) FM 4–64 permeability of wild type or cej-1+B0280.5(coRNAi). DIC image is on the left in each pair. (C,D) Embryos are at the one-cell stage except wild type stained for PGL-1, which is at the two-cell stage. Wild-type and B0280.5(RNAi) embryos are oriented with anterior (maternal DNA, polar-body end) on the left. cej-1+B0280.5(coRNAi) does not extrude polar bodies; therefore, it was not possible to determine the maternal end.