Fig. 4

a, b OA negatively regulates ovulation in the absence of sperm signal independent of G_αs signaling. The ovulation process of a virgin of fog-2 with gsa-1 RNAi (a) or fog-2; tbh-1 with gsa-1 RNAi (b) was observed at 36 h (t = 0), 48 h (t = 12), and 57 h (t = 21) after the late L4 stage. Triangle indicates vulva, red arrows mature oocytes, and yellow dotted circle a matured oocyte ovulated in the uterus or trapped inside the spermatheca. At time point 0, oocytes were stacked in both fog-2; gsa-1RNAi and fog-2; tbh-1; gsa-1RNAi (n = 12 gonads for both). c–e Representative images of fog-2 and fog-2; tbh-1 before and after mating (5 h) and OA treated. Similar images were observed from 13 animals for fog-2, 12 animals for fog-2; tbh-1 and 11 animals for fog-2; tbh-1 + OA. Red arrow: Emo. f Mating does not produce Emo when late L4 females of fog-2; tbh-1 mutants (similar images were observed from 6 animals) were mated with wild-type males. We observed 98 ovulations occurring in 25 h (~30 min/ovulation/arm). Sperm are indicated by the blue arrow). a–f Scale bar = 50 μm. g Percent unhatched embryos. fog-2 (n = 400 embryos), fog-2; tbh-1 (n = 358 embryos), fog-2; tbh-1 (OA) (n = 948 embryos) and fog-2; tbh-1 (YA) (n = 437 embryos). Two-sided t-test: p = 0.0012 (**) for fog-2 vs. fog-2; tbh-1, p = 0.0001 (***) for fog-2; tbh-1 vs. fog-2; tbh-1 + OA, p = 0.28 for fog-2 vs. fog-2; tbh-1 + OA. Boxplots show the median, mean (X), interquartile range (IQR). The upper whisker: the maxima smaller than 1.5 times IQR plus the third quartile, the lower whisker: the minima larger than 1.5 times IQR minus the first quartile. Source data are provided as a Source Data file.