Figure 10
Inhibition of autophagic flux and suppression of mitochondrial respiration by Dox-treatment in cultured cardiomyocytes. (A) Representative Western blot and densitometric quantification showing LC3B II expression at different doses of Dox (1–25 μM, 24 hours) in NRCs. GAPDH was used as a loading control. n = 6 independent experiments. (B) Inhibition of autophagic flux by 10 μM Dox (24 hours) in NRC examined by immunoblotting of LC3B II. Bafilomycin A1 (50 μg/mL) was added for 4 hours to block lysosomal degradation, and GAPDH was used as a loading control. n = 6 independent experiments. Bars represent mean ± SEM. (C) Dox treatment (1–10 μM Dox, 24 hours) dose-dependently suppress mitochondrial oxygen consumption rate (OCR) profiles in NRCs. Arrow indicates the sequential addition of oligomycin (1 µM), FCCP (4 µM), and rotenone (0.5 µM) plus antimycin A (0.5 µM). OCR profile are expressed as pMolesO2/min/µg of protein. Graph showing OCR under baseline as well as with the addition of oligomycin, FCCP, and rotenone plus antimycin A. Key parameters of the mitochondrial function, including basal and maximal respiration were dose-dependently decreased in Dox-treated NRCs. Bars represent mean ± SEM. n = 5 wells per group. P values were determined by Tukey’s post-hoc test.
