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. 2022 Jun 16;13:912648. doi: 10.3389/fendo.2022.912648

Figure 6.

Figure 6

CEFFE resists DNA damage and alleviates the senescence of KGN cells and hGCs. (A) Representative images of SA-β-gal staining. Scale bar=100 μm. (B) The senescent markers P21 and P16 and DNA damage marker γH2AX were detected via Western Blotting. (C) The protein expression of P16, P21, and γH2AX was analyzed. (D) qRT-PCR analysis of age-related gene expression in hGCs. (E) After being exposed to CPT and following treatment with/without CEFFE, γH2AX was detected via Western Blotting in KGN cells. (F) The protein expression of γH2AX was analyzed in KGN cells. (G) hGCs were identified using FSHR protein, which was specifically located in the cytoplasm of hGCs. FSHR was labeled with green fluorescence, and the nucleus was labeled with blue fluorescence. Scale bar=20 μm. (H) The DNA damaged marker γH2AX was detected in hGCs induced by CPT with the immunofluorescence method. γH2AX was labeled with green fluorescence, and the nucleus was labeled with blue fluorescence. Scale bar=20 μm. (I) Statistics of γH2AX-positive hGCs. CPT, camptothecin. Three independent experiments were conducted in each group, and data are represented as the mean ± SD, ns: P > 0.05, * P < 0.05, ** P < 0.01, *** P < 0.001, and **** P < 0.0001.