Fig. 5: Many functional germline genes are silenced in mutants with defects in forming perinuclear PRG-1 granules. | Nature Communications

Fig. 5: Many functional germline genes are silenced in mutants with defects in forming perinuclear PRG-1 granules.

From: GLH/VASA helicases promote germ granule formation to ensure the fidelity of piRNA-mediated transcriptome surveillance

Fig. 5

a A scatter plot showing the abundance of all 22G-RNAs (left) or HRDE-1 bound 22G-RNAs (right) mapped to each CSR-1 target in wild-type worms compared to indicated strains. (Left) Percentages of CSR-1 targets with twofold increased or decreased 22G-RNAs in mutants are shown. (Right) The percentage of significantly changed (adjusted P < 0.05 [see Methods for details] and twofold) CSR-1 targets are shown. Diagonal lines indicate a two-fold increase (top), no change (middle), or a twofold depletion (bottom) in the indicated mutant strains. b A scatter plot showing the mRNA vs 22G-RNA expression changes for CSR-1 targets in the indicated strain. Percentages in each quadrant indicate the proportion of CSR-1 targets with enhanced HRDE-1 targeting that fall in that quadrant. c 22G-RNA fold changes for CSR-1 targets with enhanced HRDE-1 targeting versus all other CSR-1 targets in the indicated strains versus wild-type worms. Statistical analysis was performed using a two-tailed Mann–Whitney Wilcoxon test. For all boxplots, lines display median values, boxes display first and third quartiles, and whiskers display 5th and 95th percentiles. d 22G-RNA accumulation for CSR-1 targets with enhanced HRDE-1 targeting compared between the indicated strains. Statistical analysis was performed using a two-tailed Mann–Whitney Wilcoxon test. For all boxplots, lines display median values, boxes display first and third quartiles, and whiskers display 5th and 95th percentiles. e 22G-RNAs distribution in the enhanced HRDE-1-associated 22G-RNA target ceh-49 in the indicated strains. f Photomicrographs of pachytene nuclei in adult gonads hybridized with specified single molecule fluorescent (smFISH) probes in the indicated strains. Nuclear DNA was stained with DAPI (Blue). Arrowheads indicate perinuclear mRNA foci. Arrows indicate cytoplasmic mRNA foci. Bar, 5 micrometers. g 22G-RNA enrichment in HRDE-1 versus CSR-1 IP experiments in the indicated strains for CSR-1 targets and WAGO targets. Dotted line indicates no enrichment for either Argonaute. Statistical analysis was performed using a two-tailed Mann–Whitney Wilcoxon test. For all boxplots, lines display median values, boxes display first and third quartiles, and whiskers display 5th and 95th percentiles. Distributions represent data collected across two biological replicates for CSR-1 IP and a single experiment for HRDE-1 IP.

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