DR, JLR and BR were in charge of the muscle mitochondrial extraction, mitochondrial respiration, proton leak measurements and data analysis.
We then investigated the functional properties (respiration, proton leak and ROS production) of skeletal muscle and liver mitochondria in vitro after activation/inhibition of avUCP.
MRL-MSCs displayed similar LEAK respiration, however, routine respiration and maximal electron transport capacity were greater.
Also, unaltered LEAK respiration (oligomycin-induced state 4 respiration) in combination with the detected increased parameters of maximal coupled and uncoupled respiration resulted in higher control ratios, confirming an increased efficacy of the mitochondria from the sepsis patients as compared with controls.
In contrast with the effects of in utero undernutrition on energetics in heart, adenylate-free leak respiration, fatty acid oxidative capacity, maximum oxidative phosphorylation capacity and oligomycin-induced leak respiration did not differ between U and C in liver homogenate.
Adenylate-free leak respiration, fatty acid oxidative capacity, maximum oxidative phosphorylation capacity and oligomycin-induced leak respiration were reduced in U compared with C. Calorie restriction also decreased adenylate-free leak respiration, fatty acid oxidative capacity and maximum oxidative phosphorylation capacity.
In addition, there were significant differences in maximum respiratory rate, non-mitochondrial respiration, and proton leak between RCC4-EV cells and RCC4-VHL cells (Supplementary Fig. 4e g) and the reprogramming is suppressed by YC-1 treatment (Supplementary Fig. 5a).
(B) Quantitative analysis of basal oxygen consumption, ATP production, maximal respiration and proton leak of iPSCs (n = 3, error bars represent ±SEM; **P < 0.05).
(D) Quantitative analysis of basal oxygen consumption, ATP production, maximal respiration and proton leak of NPCs (n = 3, error bars represent ±SEM; **P < 0.05).
Oxygen consumption was evaluated for cellular routine respiration, oligomycin-inhibited leak respiration, FCCP-stimulated uncoupled respiration (ETS) and rotenone/antimycin-inhibited residual respiration (ROX).
