Figure 8
mPRα was regulated by PGRMC1 during primordial folliculogenesis.
For this, 16.5 dpc ovaries were transfected with scrambled siRNA (Scra-siRNA) or siRNA against Pgrmc1 (Pgrmc1-siRNA) for 4 days, and the expression of Pgrmc2, mPRα,mPRβ and mPRγ was determined by performing qRT-PCR (A); ***denotes statistical significance at P < 0.001 between vehicle and treated ovaries (t-test, 3 independent replicates, in each replicate every treatment contained 10 ovaries). Next, Scra-siRNA- or Pgrmc1-siRNA-transfected ovaries were treated with vehicle or P4 at the same time, after 4 days of culture cell membrane protein was extracted and mPRα expression was measured by performing Western blot (B). Different letters denote statistical significance at P < 0.05 (ANOVA and post hoc test, 3 independent replicates, in each replicate every treatment contained 15 ovaries). Ovaries from 19.5 dpc were collected and subjected to immunohistochemical analysis by labeling with anti-mPRα antibodies (C). Arrows indicate labeled oocytes; scale bar = 25 μm. 16.5 dpc ovaries were treated with scrambled siRNA, scrambled siRNA plus 1 μM P4, scrambled siRNA plus 1 μM mPRα-specific agonist Org OD 02-0 (Org), or Pgrmc1 siRNA plus 1 μM Org for 6 days. Paraffinized ovary sections were labeled with hematoxylin to determine the percentage of primordial follicles (D). Different letters denote statistical significance at P < 0.05 (ANOVA and post hoc test, n = 3–5, 3 independent replicates).
