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. Author manuscript; available in PMC: 2018 Feb 1.
Published in final edited form as: J Bioenerg Biomembr. 2016 Mar 14;49(1):3–11. doi: 10.1007/s10863-016-9656-x

Figure 1. Exogenous citrate dose-dependently suppressed ROS production and depolarized mitochondria.

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Rat brain mitochondria (0.125 mg/ml) were used. The incubation medium comprised 120 mM KCl, 1 mM EGTA, 0.2 mg/ml bovine serum albumin (BSA), 4 mM KHPO4, 20 mM HEPES-KOH (pH 7.2), 5 mM succinate, 2 mM glutamate, and either 10 µM Amplex UltraRed, 4 U/ml HRP (where H2O2 emission was measured) or 2 µM Safranin O (in the membrane potential assays). A, the effect of citrate on H2O2 emission; B, the effect of citrate on the membrane potential (ΔΨm) of mitochondria; C, the dependence of the rate of H2O2 emission on the amplitude of ΔΨm; D, the dependence of the rate of H2O2 emission on the amplitude of ΔΨm in the presence of 1 mM MgCl2 added before citrate or SF6847. Insert on panel D, magnified part of the plot of citrate effect on ΔΨm and H2O2 generation. Abbreviations: cit, citrate Na+ salt; Ant.A, antimycin A 1 µg/ml; SF6847, a protonophorous uncoupler SF6847; R.F.U., relative fluorescence units. Typical curves are shown. Additions: A, B, sodium citrate as indicated; 10 nmol SF6847, 1 ng/ml Antimycin A; C, D, sodium citrate was added at 0.1, 0.4, 0.8, 1.6, 3.2, and 6.4 mM; SF6847, 2 nM each addition (16 nM, total).