Figure 2: Impact of NADH on ROS.

(A): Schematic showing the incubation system to generate NADH from PDH, with export of acetyl-CoA from mitochondria by carnitine, thus avoiding engagement of the TCA cycle. (B): NADH fluorescence in isolated mitochondria (λ_EX 340nm /λ_EM 460nm), with sequential additions of pyruvate, carnitine, ADP, and rotenone. Representative trace from 3 similar experiments. (C): pHPA fluorescent traces (individual traces in gray, average in black). Where indicated, pyruvate, carnitine, succinate, S1QEL, S3QEL and H₂O₂ were added sequentially. (D): Quantitation of H₂O₂ generation rates from traces as shown in C. Individual data points are superimposed on bars showing means ± SEM. **p<0.005 vs. baselne condition. NS: not significant.