Fig. 2

Partial mitochondrial depolarization produces a transient PINK1 response. a fluorescence timelapse microscope of HeLa cells expressing with PINK1-EGFP and mito-mCherry using the indicated CCCP treatment regimes (top). Corresponding single-cell trajectories of PINK1-EGFP fluorescence from 4 representative cells is shown for each treatment (bottom). b for the experiment described in (b), for each treatment group, cells were categorized as “no increase”, if no stabilization of PINK1-EGFP was observed post-CCCP, “transient increase” for cells where PINK1-EGFP increased post-CCCP but returned to basal levels within 2 h, and “stable increase” for cells showing a stable increase in PINK1-EGFP levels. Data is from a minimum of 3 biological repeats with ≥71 cells per condition. Error is represented as the S.E. c timelapse fluorescence images of HeLa cells expressing with PINK1-EGFP (green) and mito-mCherry (red) incubated for the indicted times with 5 μM CCCP. Scale bar represents 50 μm. d zoomed images of areas represented by the dashed boxes in (c). Scale bar represents 20 μm. Pixel intensities for line scans (position marked with thin white line) are presented below. Arrowheads indicate where line scan crosses mitochondria