Fig. 6: MVP deficiency impairs mitophagy in ECs.

A GO enrichment analysis of the downregulated DEGs in the HUVECs upon MVP knockdown. B, C MVP knockdown on mitophagy in the HUVECs treated by CCCP (10 μM) for 2 h. B Western blot analysis of autophagy protein LC3B in cytoplasmic and mitochondrial fractions of cells (n = 3, independent biological replicates). C Representative transmission electron microscope (TEM) images of the treated cells. Red arrows indicate mitophagosome vacuoles, and blue arrows indicate autolysosome vacuoles. Scale bars = 500 nm. The numbers of mitophagosome and autolysosome vacuoles per field were calculated (n = 30). D, E Spatial co-localization of mitochondrial protein COXIV (red) with LC3B (purple) in the CD31⁺ ECs (green) of carotid arteries ligated for 7 days (n = 5). Scale bar = 20 μm. F, G Spatial co-localization of mitochondrial protein COXIV (red) with LC3B (purple) in the aortic CD31⁺ ECs (green) after 4 weeks of WD feeding (n = 5). Scale bar = 20 μm. H, I Fluorescence staining of mKeima in the magnetic beads-sorted ECs of carotid arteries ligated for 7 day. Twenty cells per group (n = 20). Scale bar = 10 μm. J, K Fluorescence staining of mKeima in the magnetic beads-sorted ECs of aortas after 4 weeks of WD feeding. Twenty cells per group (n = 20). Scale bar = 10 μm. L MVP knockdown on LC3B expression in the HUVECs treated by Bafilomycin A1 (Baf-A1, 800 nM) and CCCP (n = 3, independent biological replicates). M MVP overexpression on Caspase3 and cleaved-Caspase3 expression in the HUVECs treated by cyclosporin A (CsA) (5 μM) and TNF-α (10 μg/ml) (n = 3, independent biological replicates). Data are presented as mean ± SEM. Statistical significances are assessed by Mann-Whitney test (C), two-tailed Student’s t-test (E, G, I, K), or one-way ANOVA followed by Tukey post hoc test (B, L, M). ns, no significance. Source data are provided as a Source Data file.