Fig 5. NLP-7 expression in the uv1 cells is required for the inhibition of egg-laying.

(A) Representative DIC images of wild type (upper) and transgenic animals (ufIs118) stably expressing high levels of nlp-7 genomic sequence (NLP-7 OE, lower). Arrows indicate position of the vulva. (B) Quantification of eggs in utero. Bars represent the mean ± SEM for each genotype. n≥20 for all measurements. ****p<0.0001, ANOVA with Sidak’s post hoc test. (C) Cumulative number of eggs laid over time in transgenic animals expressing the nlp-7 genomic sequence under control of a heat shock promoter (Phsp16.2::NLP-7 OE) or control animals (non-transgenic siblings). Animals were either (i) maintained continuously at 20°C or (ii) exposed to heat shock (33°C, 30 mins). *p<0.05, **p<0.01, ***p<0.001, ****p<0.0001, t-test with Holm-Sidak’s correction for multiple comparisons. Gray shading indicates the timing and duration of heat shock. (D-I) Distribution of the developmental stages of eggs laid by either wild type adult hermaphrodites (D), or animals overexpressing nlp-7 from a 3.5 kb nlp-7 promoter (E), a 2.5 kb nlp-7 promoter in which uv1 and VC fluorescence is not observed (F), a 2.5 kb nlp-7 promoter in which uv1 and VC fluorescence is observed (G), a VC neuron specific promoter (Plin-11::pes-10) (H), or a uv1 promoter (ocr-2) (I). Animals overexpressing NLP-7 from the 3.5 kb promoter (E) lay a significantly higher fraction of late-stage embryos compared with wild type (p<0.0001, Fisher’s exact test). Exclusive expression in head and tail neurons produces no significant change (F), while there is a significant shift to later developmental stages when fluorescence is observed near the vulva (G) (p<0.0001, Fisher’s exact test). uv1-specific expression also produces a significant shift (p<0.0001, Fisher’s exact test) while VC-specific expression produces no significant difference. For each genotype, eggs laid within a 30 minute time period were evaluated. See also S3 Fig.