Fig. 4: Regulatory factors act late in germline development to control fertility.

a Quantification of hatched larvae and dead eggs (brood size) following germline-specific RNAi from the L1 stage. P values were assessed by two-tailed unpaired t test comparing the brood size (hatched larvae + dead eggs) of experimental RNAi with control RNAi. n = 16, 18, 18, 15, 17, 17, 18, 17, and 18. b Timeline of temporal germline analysis showing the time and respective life stages when germline imaging and analysis were performed following egg-laying on control and RNAi plates (RNAi from the L1 stage). c Quantification of germline length of pie-1p::mCherry::his-58; sun-1p::rde-1; rde-1(mkc36) animals at the L4 stage following RNAi from the L1 stage. Data was generated from three independent experiments, and results normalized to respective controls. n = 30, 28, 30, 27, 30, 29, 27, 26, and 27. P values assessed by one-way ANOVA with no correction for multiple comparison. d DIC (left) and fluorescent micrographs (right) of pie-1p::mCherry::his-58; sun-1p::rde-1; rde-1(mkc36) germ lines at the L4 stage following control, cdc-5L and znf-622 RNAi. Blue arrow = vulva. Scale bar = 20 μm. e Heatmap showing the percentage of germ lines producing sperm, oocytes and embryos in day 1, day 2 and day 3 of adulthood following germline-specific RNAi from the L1 stage. For day 2 and day 3 adult analysis, worms were selected from day 1 sterile animals, except for the control group, and incubated for another 48 hrs or 72 hrs prior to analysis. n = 30. f Quantification of hatched larvae and dead eggs (brood size) following germline-specific RNAi from the L4 stage. n = 18, 18, 17, 18, 18 17, 18, 18 and 18. P values assessed by two-tailed unpaired t test comparing the brood size (hatched larvae + dead eggs) of experimental RNAi with control RNAi. g–j Quantification of nuclei number of PZ (g), TZ (h), pH3⁺ (i), and SYGL-1⁺ (j) of 3xflag::sygl-1; sun-1p::rde-1; rde-1(mkc36) one-day adult germ lines following RNAi treatment from the L4 stage. n = 22, 24, 23, 24, 22, 24, 24, 24 and 24. P values were assessed by one-way ANOVA with no correction for multiple comparisons. All data generated from three independent experiments. Error bars indicate SEM. Source data are provided as a Source Data file.