Figure 6

Figure 6

AAV-mediated knockdown of WASHC3 leads to defective mitochondrial respiration in human ventricular cardiomyocytes (AC16). (A) Immunoblot of WASHC3 and α-Tubulin (loading control) using AC16 cell lysate transduced with Adeno-associated virus (AAV) harboring scramble or WASHC3 shRNA (SD, n = 4; Mann–Whitney test, *p < 0.05 vs. AAV6-Scramble-shRNA). (B) Confocal images of AC16 cells stained with F-Actin (magenta) and DAPI (cyan) at 72 h after transduction (scale bar: 50 μm). (C,D)) OCR and ECAR curves in AAV-scramble shRNA or -WASHC3 shRNA transduced AC16 cells (mean ± SEM, n = 10 well per group). Correspondent quantitative analyses of parameters reveal significantly repressed of mitochondrial respiration and glycolytic activity by AAV-mediated WASHC3 knockdown. Basal respiration, Maximal respiration, non-mitochondrial respiration, ATP production, proton leak, basal glycolysis, and induced glycolysis were showed. (SD, n = 10 well per group of AC16 cells, Mann–Whitney test, *p < 0.05, **p < 0.005, ***p < 0.001 vs. Scramble shRNA).