Research Paper Volume 13, Issue 11 pp 15193—15213

TRDMT1 participates in the DNA damage repair of granulosa cells in premature ovarian failure

TRDMT1 is involved in the regulation of GCs DNA damage and apoptosis. (A) Cells with FSHR expression are shown in red. Cell nuclei are stained blue. Scale bar: 100μm. (B) DNA damage of KGN, CTX-KGN-PBS and CTX-KGN-BMSCs was detected by comet assay. Scale bar: 100μm. (C) Annexin V-APC/AAD staining and flow cytometry analysis of the apoptotic rates of cells in the WT, CTX-KGN-PBS and CTX-KGN-BMSCs groups. Quantification of the apoptosis rates is shown. (D) Immunofluorescence staining and Western blotting of the expression levels of TRDMT1 in each group. Scale bar: 10μm. (E) DNA damage of CTX-KGN transfected with GFP-NC, GFP-TRDMT1, Sh-RNA and Sh-TRDMT1 was measured by comet assay. Comparison of the tail DNA of cells in the four groups. Scale bar: 100μm. (F) Annexin V-APC/AAD staining and flow cytometry analysis of the apoptotic rates of CTX-KGN transfected with GFP-NC, GFP-TRDMT1, Sh-RNA and Sh-TRDMT1. Comparison of the percentage of apoptotic cells in the four groups. *p

Figure 2. TRDMT1 is involved in the regulation of GCs DNA damage and apoptosis. (A) Cells with FSHR expression are shown in red. Cell nuclei are stained blue. Scale bar: 100μm. (B) DNA damage of KGN, CTX-KGN-PBS and CTX-KGN-BMSCs was detected by comet assay. Scale bar: 100μm. (C) Annexin V-APC/AAD staining and flow cytometry analysis of the apoptotic rates of cells in the WT, CTX-KGN-PBS and CTX-KGN-BMSCs groups. Quantification of the apoptosis rates is shown. (D) Immunofluorescence staining and Western blotting of the expression levels of TRDMT1 in each group. Scale bar: 10μm. (E) DNA damage of CTX-KGN transfected with GFP-NC, GFP-TRDMT1, Sh-RNA and Sh-TRDMT1 was measured by comet assay. Comparison of the tail DNA of cells in the four groups. Scale bar: 100μm. (F) Annexin V-APC/AAD staining and flow cytometry analysis of the apoptotic rates of CTX-KGN transfected with GFP-NC, GFP-TRDMT1, Sh-RNA and Sh-TRDMT1. Comparison of the percentage of apoptotic cells in the four groups. *p<0.05, **P<0.01, and ***P<0.001. Statistical significance was determined using two-tailed t-tests for two groups and ANOVA for multiple comparisons. All values are means ± SD.