Fig. 3.

Effects of mitochondria-targeted MitoPQ and MitoMet on subcellular redox state. Raw264.7 cells were treated with either DTPP + or TPP + conjugated metformin, MitoPQ or unconjugated metformin for 16 h at the indicated doses. A, a representative immunoblot analysis of HIF1α and loading control GAPDH (n = 3). B, ChIP analysis was performed for HIF1α binding site on Epo promoter site and enrichment of HIF1α binding was determined (n = 3 for all groups). Data are shown as mean ± S.D. C, a representative immunoblot analysis of Calcineurin A and loading control GAPDH. D Aconitase activity assay was performed with 200 μg of total cell lysates as an indicator of oxidative stress and expressed as nmol/min/mg of protein. (means ± S.D., n = 3). E, a representative immunoblot analysis of cell extracts for PHD1, PHD2, PHD3 and loading control GAPDH following treatment with indicated doses of MitoPQ and MitoPQ control. Significance was calculated by one-way ANOVA with Tukey's multiple-comparison test, and data are presented as treatment group versus no-treatment control. In all experiments error bars represent standard deviations (*p < 0.05, **p < 0.01).