Fig. 1

The ovulatory signal activates the Hippo pathway in granulosa cells in vivo. A Levels of Hippo pathway effectors in GCs of eCG-primed mice treated with hCG on a time course. Immunoblots show 2 replicates per time point, whereas quantification was done using 4 replicates per time point. Each replicate represents the ovaries of one mouse. β-actin (ACTB) was used as the loading control. Data were normalized by the sum of all data points in a replicate and were subjected to arcsine transformation, as they are expressed as a proportion (0–1) of the whole replicate. Data were compared to 0 h hCG. One-way ANOVA (Dunnet’s post hoc test): *P ≤ 0.05, **P ≤ 0.01, ***P ≤ 0.001, ****P ≤ 0.0001. B Immunohistochemical analyses of P-LATS1(T1079) and P-YAP1(S127) were performed on ovarian sections from immature eCG-primed mice prior to or after 4 h of hCG. Negative controls were incubated without the primary antibody. Scale bars are 100 μm. C RT-qPCR analysis of Hippo pathway effectors and target genes was performed on GCs isolated from immature eCG-primed mice treated with hCG on a time course (n = 4 mice/time point, done three times). All data were normalized to the housekeeping gene Rpl19. Different letters above histograms indicate significant differences between groups. One-way ANOVA (Tukey’s post hoc test): P ≤ 0.05