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. 2022 Jun 20;23(12):6873. doi: 10.3390/ijms23126873

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© 2022 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Electron flow capacity and coupling efficiency of mitochondrial ETC in alb-SREBP-1c mitochondria. Oxidative capacity was measured in response to manipulation of the electron transport chain (ETC) in fractions of enriched mitochondria from alb-SREBP-1c vs. C57Bl6 control livers. (A) The electron flow was assayed in the uncoupled state of the mitochondrial membrane with a combination of pyruvate, malate, and succinate as substrates for oxidation using two different injection strategies. Recorded oxygen consumption rates (OCR) over the time course of the assays are shown. (B) Complex I-specific OCR as difference between stimulation of complex I with pyruvate and malate and inhibition of complex II with malonate. (C) Complex II-specific OCR as difference between stimulation of complex II with succinate and inhibition of complex I with rotenone. (D) Complex III-specific OCR after inhibition with antimycin A and (E) complex IV-specific OCR after stimulation with TMPD/ascorbate. (F) Coupling experiments were conducted by measuring OCR individually for complex I- and II-driven mitochondrial coupling in fractions of isolated liver mitochondria. (G) Basal respiration (state 2), (H) oxidative phosphorylation (state 3), (I) maximal respiratory capacity (state 3u), and (J) respiratory control ratio were calculated. Data are expressed as means (±95% CI, n = 6 per group). Statistics: Mann–Whitney test or two-way ANOVA with Tukey’s multiple comparison, * p < 0.05, ** p < 0.01, *** p < 0.001, **** p < 0.0001. Malo: Malonate, Pyr/Mal: Pyruvate and Malate, AntiA: Antimycin A, TMPD/Asc: TMPD and Ascorbate, Rot: Rotenone, Succ: Succinate, Oligo: Oligomycin, C57: C57Bl6, alb1c: alb-SREBP-1c.