Fig. 1.

The inhibitory effects of MT on autophagy reduces oxidative damage in mouse follicular GCs. Mice were injected i.p. with melatonin (15 mg/kg) or 0.5% ethanol saline once daily at 8:00 a.m. for 7 days. From day 3 to day 7, mice received an additional i.p. injection of 3-NP (50 mg/kg) or 0.5% ethanol saline at 8:00 p.m. each day. Ovaries were collected 24 h after the final injection. Immunohistochemical staining of GCs in ovarian sections was detected using anti-MAP1LC3B (A) and anti-SQSTM1 (B). Bar, 100 µm. O, oocyte; GC, granulosa cells; B, basement membrane; T, theca cells. Areas outlined in red are enlarged in lower panels. (C) Immunoblotting analysis of MAP1LC3B and SQSTM1 in ovarian GCs harvested from mice subjected to the indicated treatments. (D–F) Quantification of total MAP1LC3B expression, MAP1LC3B-II accumulation, and SQSTM1 degradation. TUBA1A served as the control for loading. Data represent mean ± S.E; n = 3. **P < 0.01; NS, not significant, P > 0.05. (G) Viability of ovarian GCs retrieved from mice with indicated administration was assessed using CCK-8 assay. Data represent mean ± S.E; n = 3 in each group. **P < 0.01; NS, not significant, P > 0.05 (For interpretation of the references to color in this figure legend, the reader is referred to the web version of this article.).