Fig. 3.
Effect of SkQ1 and C12TPP on isolated rat -liver (A) and rat-heart (B–F) mitochondria. (A) SkQ1 potentiates an oleate-induced Δψ decrease in rat-liver mitochondria. At zero time, rat-liver mitochondria (0.7 mg protein/mL) were added. Other additions, oligomycin, 1 μg/mg protein; ethanol, 2.5 μL (curve 1); SkQ1 in 2.5 μL ethanol, final concentrations 20 nM (curve 2), 40 nM (curve 3), 60 nM (curve 4), or 80 nM (curve 5); 3 × 10-6 M oleate (each addition); BSA, 0.2 mg/mL (each addition); 4 × 10-5 M 1,4-dinitrophenol (DNP). Incubation mixture, 250 mM sucrose, 5 mM MOPS-KOH (pH 7.4), 1 mM EGTA, BSA (0.1 mg/mL), 4 mM pyruvate, and 1 × 10-5 M safranin O. (B) C12TPP enhances uncoupling action of palmitate on respiration of rat heart mitochondria. Incubation medium, 250 mM sucrose, 1 mM EGTA, 10 mM MOPS-KOH (pH 7.4), oligomycin (2 μg/mL), 2 μM rotenone, 5 mM succinate, rat heart mitochondria (0.15 mg protein/mL), and, where indicated, 2.5 μM C12TPP. (C–F) SkQ1 and C12TPP effects on H2O2 formation by rat-heart mitochondria. Incubation medium, 250 mM sucrose, 1 mM EGTA, 10 mM MOPS-KOH (pH 7.4), 5 mM succinate, 2 μM Amplex Red, horseradish peroxidase (9 units), and rat-heart mitochondria (0.15 mg protein/mL). In (D), where indicated, 1 μM C12TPP was added. In (E) and (F), additions were 2 μM palmitate, 1 μM C12TPP, and 1 μM Catr.