i, quantification of average mitochondrial mass for cells exposed for 16 h to the indicated concentrations of CCCP. h fluorescence images of SH-SY5Y cells stained with MitoTraker Green FM (green) and Hoechst 33342 16 h post-CCCP. Data from 4 representative cells is shown. g CCCP treatment regimes (top) and corresponding single-cell trajectories of TMRM fluorescence in SH-SY5Y cells (bottom). The average number of mtDNA puncta were quantified for 20 cells per condition and presented in ( f) normalized to the number of puncta in control cells. e + f HeLa cells expressing EYFP-Parkin (green) were incubated with the indicated concentrations of CCCP for 24 h, fixed, and stained with DAPI (blue) and antibodies to detect mtDNA nucleoids (red), then imaged by confocal microscopy. For ( c) and ( d), error is represented as the standard error (S.E.). Data is presented as an average of 3 biological repeats with a minimum of 18 cells per condition. d average TMRM staining for cells 120 min post-CCCP treatment. c population average trajectories of TMRM fluorescence for cells depicted in ( b).

Trajectories for each cell are represented in a different colour. Data from 4 representative cells is shown for each treatment. b CCCP treatment regimes (top) and corresponding single-cell trajectories of TMRM fluorescence in HeLa cells (bottom). a fluorescent timelapse microscopy images of TMRM-stained HeLa cells at the indicated times post 10 or 5 μM CCCP treatment. Complete and partial depolarization of mitochondria through titration of CCCP.

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