Fig. 4

Dancr^−/− mice display impaired fertility and POI phenotypes. A, The Dancr knockout mice was identified by qPCR. B, The ISH results illustrated Dancr was specifically expressed in normal ovarian GCs and knocked out in GCs of Dancr^−/− mice. Scale bars: 200 µm (overview), 50 µm (zoom-in). C, The live birth number of Dancr^+/+ and Dancr^−/− mice per litter. D, Left, Four patterns of estrous cycles observed in the study: I, normal cycles; II, disordered cycles with a prolonged diestrus and normal estrus; III, disordered cycles with prolonged estrus and diestrus; IV, disordered cycles with a prolonged estrus and normal diestrus. The x axis represents the number of observing days and y axis represents the cycle day in proestrus or estrus (P-E) and metestrus or diestrus (M-D). Right, The ratio of the mice with normal or disordered estrous cycle. E, Serum AMH, FSH and E₂ levels in 8-weeks Dancr^+/+ and Dancr^−/− mice. F, Left, HE staining sections of mouse ovaries, arrow indicates atresia follicle. Right, Total number of mouse follicles in all serial sections and the proportion of follicles at different stages. Scale bar: 200 µm. Pmd represents primordial follicle; Antral represents antral follicle; Atresia represents atretic follicle. G, SA-β-Gal staining of mice primary ovarian granulosa cells and quantify analysis in Dancr^−/− and Dancr.^+/+ mice. Scale bar: 50 μm. H, Protein levels of p53 and p21 were detected using Western blot analysis. *p < 0.05, ** p < 0.01, *** p < 0.001