Immunohistochemistry analysis was performed on ovary sections to identify the oocytes. Briefly, the ovaries from E13.5, PD1, and PD8 were fixed with 4% paraformaldehyde for 24 hours, then washed with PBS, and stored in 70% ethanol. The samples were embedded in paraffin, and 8-μm sections were prepared. After deparaffinization and rehydration through a graded ethanol series, the slides were incubated with 5% BSA for 1 hour at room temperature and incubated with anti-MVH (1:5000 dilution; Abcam #ab27591) antibody overnight at 4°C. Quantification of ovarian follicles was performed as previously described (53). Briefly, MVH-positive oocyte nuclei with characteristic surrounding granulosa cell layers were counted for each section. Counts were added to calculate the total number of oocytes per ovary. For each genotype, three to six ovaries were analyzed.

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