Figure 2.

Overexpression of MARCH5 reduces cardiotoxicity induced H₂O₂. (A) H9C2 cells were transfected with MARCH5‐cDNA for 24 h, and the expression level of MARCH5 was detected by Western blotting. (B) H9C2 cells were exposed to 100 μM H₂O₂ for another 24 h, and mitochondrial morphology was stained with MitoTracker Red and observed using a laser‐scanning confocal microscope, (C) shown the percentage of cells undergoing mitochondrial fission. Apoptotic cells were detected by TUNEL assay (D) and the percentage of apoptotic cells shown in (E). Autophagy flux was assessed with transduced Ad‐RFP‐GFP tandem‐tagged LC3. Autophagy flux was observed using a laser‐scanning confocal microscope (F) and (G) shown the numbers of autolysosomes and autophagosomes. LC3 protein was detected by Western blotting (H) and densitometry (I). All of the data were expressed as the mean ± SEM of three independent experiments. *P < .05, **P < .01, ***P < .001