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Figure 4 Anti-erbB2 reduces mitochondrial membrane potential adenosine triphosphate (ATP) concentration and redox capacity in neonatal rat ventricular cardiomyocytes. Neonatal rat ventricular cardiomyocytes were stained with a fluorescent probe selective for metabolically active mitochondria Mitotracker red. (a) Results of flow cytometry of cells stained with Mitotracker red dye. Anti-erbB2 treatment resulted in decreased red fluorescence (76.85 ± 2.4 vs. 51.1 ± 0.072, *p < 0.05, n = 4 independent experiments). (b) ATP content was measured using luciferin-luciferase bioluminescence. There was a 35% decrease in ATP content of anti-erbB2-treated cells at 36 h compared to untreated cells. Results, expressed as relative light units, are from three independent experiments with six replicates/condition (1,297 ± 112 vs. 847 ± 75, *p < 0.01). (c) Reduction of the tetrazolium salt mitochondrial reductase activity (MTT) to a corresponding formazan is an indicator of redox capacity. Redox capacity was quantified using a spectrophotometer at a wavelength of 590. Reported results represent pooled data from nine independent experiments with three or more replicates, for each condition reported, in each experiment (1.0 vs. 0.804, *p < 0.01: normalized data pooled from nine independent experiments, three replicates/condition). Ab = antibody; M1% = percent positive of gated cells.
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