ALDH2 Activity Reduces Mitochondrial Oxygen Reserve Capacity in Endothelial Cells and Induces Senescence Properties

Figure 5

ALDH2 silencing is associated with mitochondrial dysfunction. (a) OCR was assessed by a Seahorse XF24 cell culture microplate in siCTR and siALDH2 ECs that were harvested and seeded 24 h posttransfection in XF24 cell culture plates at a density of 3 × 10⁴ cells/well. Where indicated (arrows), oligomycin (O) (1 μg × ml⁻¹), FCCP (F) (0.2–0.4 μM), rotenone (R) (1 μM), and antimycin A (AA) (1 μM) were added. Data are representative of three experiments. (b) Basal OCR, proton leak, ATP-linked OCR, maximal OCR, and reserve capacity in siALDH2 ECs exposed to 2% FBS for 24 h. The means ± SEM of each parameter are shown. and vs. siCTR. (c) Western blot analysis of OXPHOS representative complexes detected by the OXPHOS antibody cocktail kit in siALDH2 (two clones A and B) or siCTR cultured in 2% FBS for 24 h. Knockdown efficiency was verified with an ALDH2 antibody. Actin was used as a loading control. Blots are representative of 3 with similar results.

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