Fig. 4

ESCs-sEVs promoted the proliferation and inhibited the apoptosis of ovary granulosa cells. A Morphology and identification of mouse ovarian granulosa cells. A (a) Granulosa cells adhered to the wall after 24 h (× 100). A (b) Granulosa cells proliferated significantly after 72 h (× 100). A (c) The phenotype of granulosa cells is shown by H&E staining (× 200), bars = 50 μm. A (d–f) Identification of ovarian granulosa cells by FSHR immunofluorescence (× 200). Bars = 100 μm. B ESCs-sEVs promoted the proliferation of granulosa cells, as detected by EdU tests; the cell nuclei are stained blue, and the cells with proliferation activity are stained green. C The proliferation ratio of granulosa cells was analyzed using EdU tests. D The cell proliferation curve of granulosa cells was measured by CCK-8 from day 1 to day 5. E The expression of FSHR, a protein related to apoptosis and the PI3K/AKT signaling pathway, was measured in granulosa cells by Western blotting analysis. Data are presented as the mean ± SD. *P < 0.05, **P < 0.01, ***P < 0.001