Deubiquitinating enzyme USP30 negatively regulates mitophagy and accelerates myocardial cell senescence through antagonism of Parkin

摘要

Myocardial cells were treated with d-gal and Mito-TEMPO. A Representative images and quantitative analysis of cell senescence detected by SA-β-gal staining (×200). B Immunoblots of p53, p21, and p16 expression in myocardial cells and densitometric quantification of the relative band intensity. C Representative images and quantitative analysis of total ROS in myocardial cells assessed by DCFH-DA immunofluorescence assay (×400). D Representative images and quantitative analysis of mitochondrial ROS in myocardial cells examined by DHR123 immunofluorescence assay (×400). E, F MMP in myocardial cells evaluated by JC-1 immunofluorescence (E, ×200) and TMRM (F, ×400) assays. *P < 0.05 compared with control myocardial cells. #P < 0.01 compared with the myocardial cells with d-gal treatment. Measurement data are expressed as mean ± standard deviation. Comparisons among multiple groups are analyzed by ANOVA with Tukey’s post hoc test. The experiment is repeated three times.