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. 2020 Oct 7;37:101750. doi: 10.1016/j.redox.2020.101750

Fig. 2.

Fig. 2

Lack of USF2 alters mitochondrial morphology and function. (A) Control, ΔUSF2 and ΔUSF2+USF2 MEFs cells were subjected to electron microscopy. Representative images of a cross-section showing the mitochondria (arrows), are presented. Original magnification, 6300 ×. Scale bar 1 μm. (B) Control, ΔUSF2 and ΔUSF2+USF2 cells were stained with TMRE and analyzed with the Operetta high-content imaging system. *significant difference control vs ΔUSF2 cells, **ΔUSF2 cells vs ΔUSF2+USF2 cells, p ≤ 0.05. (C) Representative fluorescent images of TMRE stained cells. (D) The oxygen consumption rate (OCR) of control and ΔUSF2 cells was measured under basal conditions and after the sequential addition of 1 μM oligomycin, 1 μM FCCP, and 0.5 μM rotenone/antimycin to allow calculation of basal respiration, ATP production, proton leak, and spare respiratory capacity (SRC). *significant difference, p ≤ 0.05. (E) Loss of USF2 increases ROS levels. Control, ΔUSF2 and ΔUSF2+USF2 MEFs were stained with CellROX® and the fluorescence intensity was analyzed and quantified with the Operetta high-content imaging system. *significant difference control vs ΔUSF2 cells, **ΔUSF2 cells vs ΔUSF2+USF2 cells, p ≤ 0.05. (F) Representative images of cells displaying CellROX® fluorescence.