Research Paper Volume 13, Issue 18 pp 22444—22458

SUV39H2/KMT1B Inhibits the cardiomyocyte senescence phenotype by down-regulating BTG2/PC3

Knockdown SUV39H2 aggravate senescence in H2O2-treated H9C2 cells. (A) H9C2 were infected with lenti-shRNA or lenti-shSUV39H2-1 or lenti-SUV39H2-2 in 50 μM H2O2 for 48 hours, then protein levels of SUV39H2 were detected by western blot. (B) H9C2 were as described above for 48 hours, and β-galactosidase staining was performed. The positive cells are shown in blue. (C–H) H9C2 were cultured with basic medium or 50 μM H2O2 supplemented with or without the lenti-sh-SUV39H2 for 48 hours. (C–E) The protein and mRNA expression of p53, p21, and SUV39H2 were performed by Western blot and qRT-PCR, and ROS generation was detected. (F) Detection of γ-H2AX expression in each group by immunofluorescence. (G) JC-1 monomer images were shown. The green fluorescence represents JC-1 monomers in 530 nm, whereas red fluorescence represents JC-1 aggregates in 590 nm. (H) Electron microscopic images of mitochondrial. All the experiments have been repeated independently at least 3 times. Scale bars, 200 μm.*P **P ***P

Figure 2. Knockdown SUV39H2 aggravate senescence in H2O2-treated H9C2 cells. (A) H9C2 were infected with lenti-shRNA or lenti-shSUV39H2-1 or lenti-SUV39H2-2 in 50 μM H2O2 for 48 hours, then protein levels of SUV39H2 were detected by western blot. (B) H9C2 were as described above for 48 hours, and β-galactosidase staining was performed. The positive cells are shown in blue. (CH) H9C2 were cultured with basic medium or 50 μM H2O2 supplemented with or without the lenti-sh-SUV39H2 for 48 hours. (CE) The protein and mRNA expression of p53, p21, and SUV39H2 were performed by Western blot and qRT-PCR, and ROS generation was detected. (F) Detection of γ-H2AX expression in each group by immunofluorescence. (G) JC-1 monomer images were shown. The green fluorescence represents JC-1 monomers in 530 nm, whereas red fluorescence represents JC-1 aggregates in 590 nm. (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.