Fig 1: The mRNA levels and protein expressions of genes regulated by PGR were downregulated in the mice of PPOS group(A–D) The relative mRNA levels of HIF (A), ADAMTS-1 (B), VEGF-A (C), and EDN2 (D) in the control group (n = 6, 8, 11, 10 mice in A–D) and PPOS groups (n = 9, 10, 10, 9 mice in A–D).(E–H) The immunofluorescence images of HIF (E), ADAMTS-1 (F), VEGF-A (G), and EDN2 (H) expression in the ovaries (n = 3–5 mice in each group). The preovulatory follicles indicated by asterisks in the upper panel were enlarged in the lower panel. Scale bar in the upper panel, 500 μm. Scale bar in the lower panel, 200 μm.(I–L) The relative fluorescence intensity of HIF (I), ADAMTS-1 (J), VEGF-A (K), and EDN2 (L) within control and PPOS groups analyzed by ImageJ software (n = 6–8 preovulatory follicles within 3–5 mice in each group).Statistical analysis was performed by Mann-Whitney test in A–D and I–L. Data are represented as mean ± SEM. ns, no significance (p ≥ 0.05). ∗p < 0.05. ∗∗p < 0.01. ∗∗∗p < 0.001.
Fig 2: The decreased mRNA levels of progesterone synthesis enzymes and PGR expression in the mice of PPOS group(A–D) Star (A, n = 9–10 mice in each group), Cyp11a (B, n = 9–10 mice in each group), Hsd3B2 (C, n = 10 mice in each group), and Cyp19a (D, n = 7–10 mice in each group) relative mRNA levels in the PPOS and control groups at 4 h after hCG injection.(E) Immunofluorescence images of PGR expression on the preovulatory follicles in the PPOS and control groups at 6 h after hCG injection (n = 3–5 mice in each group). Scale bar, 50 μm.(F) The relative PGR fluorescence intensity within control and PPOS groups analyzed by ImageJ software (n = 6 preovulatory follicles within 3–5 mice in each group).(G) PGR relative mRNA expression levels at 4 h after hCG injection within two groups (n = 7–9 mice in each group).Statistical analysis was performed by Mann-Whitney test in A–D, F-G. Data are represented as mean ± SEM. ns, no significance (p ≥ 0.05). ∗p < 0.05, ∗∗p < 0.01.
Fig 3: Effect of MnPP on PR-associated protein expression. Representative western blots (A and B) show expression of PR-B, EREG, P27, and SP1 in epithelial cell lysates from controls (lanes 1-3) and MnPP (lanes 4-6) animals at PND 30 and PND 120, respectively. Bar graphs show changes in the mean (±SEM) of respective protein expression levels for PR-B (C), EREG (D), Sp1 (E), and p27 (F), at PND 30 (left) and PND 120 (right). Animal numbers (N) are shown within the bars. Data was analyzed by Student t-test. *P < .05; **P < .01.
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