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This study investigates the role of ANT in proton leak in muscle and liver mitochondria from WT and Grx2-/- mice, along with UCP3 protein levels using immunoblot analysis. Results provide insights into mitochondrial function and proton leak regulation.
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Supplemental Figure 5 p<0.05 p=0.05 a b p<0.05 p<0.05 p<0.05 p<0.05 c Muscle d Liver + - + - UCP3 p<0.01 UCP3 SDH p<0.05 Figure S5: A) Contribution of ANT to proton leak in liver mitochondria from WT and Grx2-/- mice. Following a measure of state 3 respiration, mitochondria were sequentially treated with oligomycin (Oligo; 2.5 μg/mL), carboxyatractyloside (CAT; 5 μM), and antimycin A (4 μM). Oxygen consumption rate (OCR) values were corrected for background (antimycin A) respiration. n=4, mean±SEM, 2-way ANOVA with Fisher’s posthoc test. B) Contribution of ANT to proton leak in muscle mitochondria from WT and Grx2-/- mice. Following a measure of state 3 respiration, mitochondria were sequentially treated with oligomycin (Oligo; 2.5 μg/mL), carboxyatractyloside (CAT; 5 μM), and antimycin A (4 μM). OCR values were corrected for respiration not associated with the electron transport chain (antimycin A). C) Immunoblot analysis of UCP3 protein levels in mitochondria collected from WT and UCP3-/- mice. D) Contribution of ANT to proton leak in muscle mitochondria from WT and UCP3-/- mice. Following a measure of state 3 respiration, mitochondria were sequentially treated with oligomycin (Oligo; 2.5 μg/mL), carboxyatractyloside (CAT; 5 μM), and antimycin A (4 μM). OCR values were corrected for respiration not associated with the electron transport chain (antimycin A)Student t-test, n=4, mean±SEM.
