For imaging mitochondrial mass or morphology independent of membrane potential [21 (link)], live cells were stained by MitoTracker Green (100 nmol·L− 1, Invitrogen) at 37 °C for 30 min or transfected with MitoRFP reporter (Addgene) for 48 h, with nuclei staining by Hochest 33,342 (Sigma). Mitochondrial structure was observed by transmission electron microscopy [22 (link)]. Cells were treated with JC-1 (10 μg/ml, Sigma) at 37 °C for 20 min, while red fluorescence intensity reflecting Δψm was assessed with a fluorescence microscope. Mitochondrial complex I activity was determined by Mitochondrial Complex I Activity Assay Kit (Sigma).
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