Fig. 3: Granulosa cell secreted VEGFA governs the ovarian angiogenesis.
a In situ hybridization demonstrating granulosa cells (arrows) as the primary cells of VEGFA secretion in ovaries. Three experiments were repeated independently with similar results. b Schematic representation of the strategy of Vegfa deletion in GCs. Exon 3-7 deletion via Foxl2-Cre-mediated recombination in granulosa cells within GC;Vegfafl/fl (vko) ovaries. c Validation of Vegfa deletion in GCs of vko females. Three experiments were repeated independently with similar results. d Histological analysis showing a significant suppression of ovarian and follicle development in the vko females compared to nocre ovaries at 23 dpp and 6 months. No ovulated follicles and many empty follicle-like structures (arrows) were observed in vko ovaries at 6 months. e Follicle counting results in nocre ovaries at 23 dpp and 6 months, showing a dramatically decreased number of growing follicles at 23 dpp (left) and a failure of ovulated follicle formation at 6 months (right) in vko ovaries (23dpp: n = 4 mice, 6 M: n = 3 mice). f Immunostaining of endothelium revealing the failure of follicle blood vessel formation (arrows) in vko ovaries (green: PODXL, gray: Hoechst). g Quantification of relative vascular density in vko ovaries at both 23 dpp and 6 months (n = 3 mice). h Histological analysis of ovaries before and after PMSG treatment, showing a failure of PMSG response with no markedly changes of in both ovarian size and follicle developmental profiles in vko ovaries after PMSG treatment. i Maximum follicle diameter in 23 dpp ovaries before and after PMSG treatment, showing a significantly increase of follicle diameter in nocre ovaries, but the size of largest follicles had no changes in vko ovaries (n = 10 follicles from 3 mice). Data are presented as the means ± SD. Data were analyzed by 2-tailed unpaired Student’s t-test without adjustments; *** P < 0.001, ** P < 0.01, n.s. P ≥ 0.05. Scale bars: 200 μm (a), 100 μm (d and f), 500 μm (h). Source data are provided as a Source Data file including exact P values.
