Figure 6.

Mitochondrial Transfer Reduces ROS, Augments ATP, and Preserves Mitochondrial Biogenesis

(A) iCMs treated with L-EVs at 24 hours following DOX injury showed reduced cytoplasmic (top, CellROX Deep Red) and mitochondrial (bottom, CellROX Green) ROS production (n = 6). (B) Following DOX injury, iCMs treated with L-EVs demonstrate greater ATP production per iCM (n = 4). (C, left) Representative western blot showing Peroxisome proliferator-activated receptor-gamma coactivator 1 (PGC1)-alpha expression in iCMs. (C, right) ImageJ analysis of relative PGC1-alpha expression in 3 iCM groups following DOX injury. iCMs treated with L-EVs show greater expression than both no DOX and DOX-injured iCMs (n = 4). (D) 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolim bromide 48-hour viability assay showing MPP+ iodide attenuates the therapeutic effect of L-EVs. Red blood cell (RBC)–derived L-EVs lack mitochondria and show no therapeutic effect. Dexrazoxane (DZR) serves as positive control (n = 9). ∗P < 0.05, ∗∗P < 0.01, and ∗∗∗P < 0.001 by 1-way analysis of variance or 2-way analysis of variance with repeated measures followed by Tukey's test. Abbreviations as in Figures 1 and 3.