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. Author manuscript; available in PMC: 2016 Sep 1.
Published in final edited form as: Free Radic Biol Med. 2015 Jun 6;86:250–258. doi: 10.1016/j.freeradbiomed.2015.05.032

Fig. 6.

Fig. 6

MitoSOX impairs both complex I- and complex II-dependent ADP-stimulated respiration. A. Primary rat cortical neurons were treated with vehicle control (con, filled circles) or MitoSOX (10 μM, open circles) after three baseline O2 consumption rate (OCR) measurements (first arrow). Neurons were permeabilized by saponin (sap, 25 μg/ml) in the presence of EGTA (5 mM, second arrow). Pyruvate and malate (P/M, 5 mM each), ADP (1 mM), and excess K2PHO4 (3.6 mM for 4 mM final) were co-injected with saponin to measure complex I-dependent ADP-stimulated respiration. Purified cytochrome c (cyt c, 100 μM) was injected at the third arrow, followed by antimycin A (AA, 1 μM) at the fourth arrow. B. The same experiment as depicted in A, but with the complex II substrate succinate (S, 5 mM) and the complex I inhibitor rotenone (R, 0.5 μM) added in place of complex I substrates pyruvate and malate. Results in A and B are mean ± SD (n=3 wells) and are representative of two independent experiments. OCR is baseline-normalized to the point immediately prior to the first addition.