Research Paper Volume 13, Issue 18 pp 22444—22458

Figure 3. Overexpression SUV39H2 alleviate senescence in H₂O₂-treated H9C2 cells. (A) The SUV39H2-overexpression plasmid was constructed and verified by western blotting. (B–F) H9C2 were cultured with basic medium or 50 μM H2O2 supplemented with or without the lenti-ov-SUV39H2 for 48 hours. (B) The image shows that each group was stained with β-galactosidase, and the percentage of total cells stained with β-galactosidase are blue. (C) Western blotting was used to detect the expression of p53, p21 and FLAG. (D) Flow detection of ROS yields in each group. (E) The expression of p21 and p53 was quantified using RT-PCR. (F) Detection of γ-H2AX expression in each group using immunofluorescence. (G) Overexpression of SUV39H2 in H9C2 cells stained with JC-1 at 48 hours after treatment with H₂O₂. The green dye represents JC-1 monomers in cytoplasm while in red represents JC-1 aggregates in nucleus. (H) Electron microscopic images of mitochondrial. All the experiments have been repeated independently at least 3 times. Scale bars, 200 μm. ^*P < 0.05, ^**P < 0.01, ^***P < 0.005 when two groups were compared as indicated, or compared to the corresponding control.