Fig. 3

GCN5L1 deficiency restores mitochondrial oxidative phosphorylation. A and B CFs were treated with or without Ang II after transfection with an empty vector or sh-GCN5L1. The experimental program (A) and statistical analysis (B) showing the extracellular acidification rate (ECAR) of CFs as measured with a Seahorse XFe96 Extracellular Flux Analyzer (n ≥ 10/group). C and D CFs were treated with or without Ang II after transfection with an empty vector or LV-GCN5L1. The experimental program (C) and statistical analysis (D) of the extracellular acidification rate (ECAR) of the CFs as measured with a Seahorse XFe96 Extracellular Flux Analyzer (n ≥ 11/group). E and F CFs were treated with or without Ang II after transfection with an empty vector or sh-GCN5L1. The experimental program (E) and statistical analysis (F) of the oxygen consumption rate (OCR) of CFs as measured with a Seahorse XFe96 Extracellular Flux Analyzer (n ≥ 10/group). G and H CFs were treated with or without Ang II after transfection with an empty vector or LV-GCN5L1. The experimental program (G) and statistical analysis (H) of the oxygen consumption rate (OCR) of the CFs as measured with a Seahorse XFe96 Extracellular Flux Analyzer (n ≥ 10/group). I Schematic diagram showing isotope-tracing experiments. J, K, L CFs were induced differentiation with or without Ang II. [U-¹³C₆] glucose was added into culture media on 24 h for 30 min. Then, tracing analysis from U-¹³C₆ glucose was performed. Intracellular abundance of pyruvate (M + 3) (J). Intracellular abundance of lactate (M + 3) (K). Intracellular abundance of citrate (M + 2) (L). Data are the means ± SEM of at least 6 independent experiments. The results are presented as fold increases relative to vector control. ^*p < 0.05, ^**p < 0.01. M and N the experimental program (M) and statistical analysis (N) of the effect of the extracellular acidification rate (ECAR) on in CFs pre-treated with UK5099 for 1 h then cultured with or without Ang II, which was measured with a Seahorse XFe96 Extracellular Flux Analyzer (n ≥ 11/group). O and P CFs were pre-treated with methyl pyruvate for 1 h then cultured with or without Ang II. The experimental program (O) and statistical analysis (P) of the effect of the oxygen consumption rate (OCR) on these cells as measured with a Seahorse XFe96 Extracellular Flux Analyzer (n = 12/group). The data are shown as the mean ± SEM. P values were calculated by one-way ANOVA. ^*p < 0.05, ^**p < 0.01