Fig. 4: PTV activated PINK1-PARK2 dependent mitophagy.

a Representative western blots for the detection of ATG7, ATG12–ATG5 conjugate, MAP1LC3B-II, and SQSTM1 after infection showed that Atg7 was successfully knocked down and mitophagy was effectively inhibited in Atg7 silencing group. b Silencing Atg7 before PTV treatment significantly reduced proliferative activity compared with PTV alone group. c 3-MA (2 mM) was added to inhibit mitophagy before 0.5 μM PTV treatment. CCK-8 assay showed that the proliferative activity in 3-MA + PTV group reduced significantly compared with PTV alone group. (d–f) PTV treatment (0, 0.1, 0.5, or 1.0 μM) for 24 h increased PINK1 accumulation and PARK2 recruitment in mitochondrial membrane in dose-dependent manner. g Mitotracker Deep Red was used to mark mitochondria and immunostain was employed to mark PARK2 in atherosclerotic mice EPCs. Merged images and Pearson’s overlap coefficient analysis indicated that PARK2 mostly localized on mitochondria in PTV treatment EPCs related to HFD group, scale bar: 10 μm. h Atherosclerotic mice EPCs were co-immunostained for MAP1LC3B and PARK2. Overlap of PARK2 with MAP1LC3B were observed in PTV treatment EPCs in comparison to HFD group and Pearson’s overlap coefficient was employed to analyze the co-localization, scale bar: 10 μm. (n = 10 cells per group, EPCs were isolated from ApoE^−/− mice fed with high-fat diet for 8 weeks, cells were isolated from 3 mice for 1 experiment and 3 independent experiments were performed, mean ± SD, *P < 0.05, **P < 0.01).