Fig. 5
Effects of Gdf9 variants on antral follicle development after PMSG stimulation. A Appearance of Gdf9wt/wt, Gdf9S415T/S415T and Gdf9Q308X/S415T ovaries 48 hours after PMSG stimulation as viewed through the stereoscope. B Appearance of cumulus-oocyte complex collected 12 hours after HCG injection. C H&E staining of Gdf9wt/wt (n=3), Gdf9S415T/S415T (n=3) and Gdf9Q308X/S415T (n=3) ovaries 48 hours after of PMSG stimulation. The images shown were the sections where the largest follicle of each ovary was located. The follicle diameter, average thickness of the mural granulosa cell layer and cumulus cell layer were calculated for the largest follicle. *P < 0.05. D Ki-67 immunostaining in Gdf9wt/wt (n=3), Gdf9S415T/S415T (n=3) and Gdf9Q308X/S415T (n=3) ovaries. The ki67 positivity rates of cumulus cells and mural granulosa cells in large follicles, and the ki67 positivity rates of granulosa cells in small follicles were calculated separately. *P < 0.05. E TUNEL (red) and DAPI (blue) immunofluorescence in Gdf9wt/wt (n=3), Gdf9S415T/S415T (n=3) and Gdf9Q308X/S415T (n=3) ovaries. The dotted lines outline the boundaries of granulosa cells. Arrows indicate TUNEL-positive cells. TUNEL positivity rates of granulosa cells in each antral follicle were calculated