Loss of function of factors involved in mRNA processing, transport, and translation reduces the levels of mRNA and IFE-1 partitioning into PGL granules. (A–D) Compared with control(RNAi)-treated atg-3 mutant embryos (A), the number of poly(A) mRNA-positive puncta and also the ratio of PGL granules positive for poly(A) mRNA in somatic cells is significantly decreased in atg-3 mutant embryos with simultaneous ruvb-1(RNAi) (B) and pab-1(RNAi) (C) at 26°C. Embryos at the ∼100–200 cell stage are shown in A–C. The germ precursor cells (Z2, Z3) are highlighted in dashed circles. (D and E) Quantification of the number of somatic poly(A) mRNA puncta (D) and % of PGL granules positive for poly(A) mRNA per focal plane in the indicated genotypes at 26°C (E). Data are shown as mean ± SEM (n = 5; n refers to five images from the corresponding five embryos analyzed for each genotype). Two-tailed, unpaired t test results: ***P < 0.001. (F–J) IFE-1::GFP granules are less in number and weaker in intensity in pab-1(RNAi) (G) and pab-1(RNAi); atg-3(bp412) (I) embryos at 26°C, compared with control(RNAi)-treated WT embryos (F) and atg-3(bp412) embryos (H), respectively. J shows the quantification of the number of IFE-1::GFP granules in the indicated genotypes. Data are shown as mean ± SEM (n = 3; n refers to the number of embryos analyzed for each genotype). Two-tailed, unpaired t test results: **P < 0.01, ***P < 0.001. Maximum-intensity projections of Z-stack confocal images of comma-stage embryos are shown in F–I. Scale bars: 5 µm for A–C and F–I; 2 µm for enlarged images in A–C.