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. 2022 May 3;13:2403. doi: 10.1038/s41467-022-30169-y

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© The Author(s) 2022

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 4 — a Effects of light-induced complex I ROS on (a) complex I and (b) complex II respiration. Wild-type and nuo-1::Supernova isolated mitochondria were exposed to light (GYX, 7.8 mW/mm²) for 0 or 10 min. State 3, State 4 respiration, and the RCR were measured using (a) complex I- and (b) complex II-linked substrates. Data are mean ± SD. N = 3 independent mitochondrial preparations, ns = not significant, *p < 0.05, **p < 0.01 (Two-way ANOVA, Tukey’s multiple comparisons). c Schematic diagram of the mitochondrial electron transport chain. Complex I transfers electrons to coenzyme Q which will ultimately reduce oxygen to water. Complex I activity was assessed via oxygen consumption, total activity (NADH to Q) and NADH dehydrogenase activity (NADH to hexaammineruthenium). d Effects of light-induced complex I ROS on complex II enzyme activity. Isolated freeze-thawed mitochondria were exposed to light (7.8 mW/mm²) for 0, 5, and 10 min. Complex II activity was measured as malonate-sensitive succinate oxidation rate of DCPIP reduction. Data are mean ± SD. N = 5 independent mitochondrial preparations, ns=not significant (Two-way ANOVA, Tukey’s multiple comparisons). e Effects of light-induced complex I ROS on complex I total activity. Isolated freeze-thawed mitochondria were exposed to light (7.8 mW/mm²) for 0, 5, and 10 min. Total complex I activity was measured as the rotenone-sensitive rate of NADH oxidation. Data are mean ± SD. N = 6 independent mitochondrial preparations, **p < 0.01 (Two-way ANOVA, Tukey’s multiple comparisons). f Effects of light-induced complex I ROS on complex I NADH dehydrogenase activity. Isolated freeze-thawed mitochondria were exposed to light (7.8 mW/mm²) for 0, 5, and 10 min. NADH dehydrogenase activity of complex I was measured as the hexaammineruthenium (HAR)-dependent rate of NADH oxidation. Data are mean ± SD. N = 4 independent mitochondrial preparations, ns = not significant (Two-way ANOVA, Tukey’s multiple comparisons). g NAC reverses effects of light-induced complex I ROS on complex I total activity. Isolated freeze-thawed mitochondria were exposed to light (7.8 mW/mm²) for 0, 5, and 10 min. Total complex I activity was measured in the presence of 2.5 mM NAC. Note: NAC was not present during light treatment. Data are mean ± SD. N = 4 independent mitochondrial preparations, **p < 0.01 (Two-way ANOVA, Tukey’s multiple comparisons).