Fig 2: The interactions of CHK2 with FoxK or FoxO3a. (A) The cell lysates were processed for coimmunoprecipitation with anti-CHK2, followed by probing with anti-FoxO3a, FoxK and FoxO4. IP, immunoprecipitation. (B) GCs transfected with CHK2 siRNA or scrambled control siRNA for 24 h/48 h were cultured for 24 h and then the cells were rinsed with PBS. The expression of CHK2 was determined by qPCR and the expression of CHK2 protein was determined by Western blot. Bars with different superscripts are statistically different (p < 0.05). (C) GCs transfected with CHK2 siRNA for 24 h were cultured in media containing ACNU. 24 h later, the cells were rinsed with PBS and the expressions of p53, p-p53, Rad51, CDKN1A, CCND1, LC3B and Atg7 were determined by Western blot. Bars with different superscripts are statistically different (p < 0.05).

Fig 3: Reduced DNA damage after FSH treatment in ASWFs. (A) GCs from ASWFs were treated with 0.01 IU/mL FSH in culture. Cell viability using the CCK-8 assay in GCs from ASWFs treated with FSH in culture. Values are the means ± SEM (n = 3). Bars with different superscripts are statistically different (p < 0.05). (B) Relative expression of proteins related to DNA damage in GCs from ASWFs treated with FSH. Bars with different superscripts are statistically different (p < 0.05). (C) ASWFs were treated with 0.1 IU/mL FSH for 72 h. Representative morphology and the expression of FSHR of ASWFs treated with FSH in culture. Scale bar in H&E staining: 50 µm; Scale bar in IF staining: 20 µm. GL: Granulosa layer; TL: Theca layer. (D) Relative expression of ?H2AX, FSHR, caspase3, PCNA, Rad51 and CHK2 proteins in cultured ASWFs treated with FSH. Bars with different superscripts are statistically different (p < 0.05).