Fig. 7.

A‐769662‐treated mitochondria are fragmented and accumulate in the perinuclear region. (A) Representative confocal microscopy images from uninduced and induced (6 days) U2OS Polɣ^D1135A cells either untreated or treated with 100 μm A‐769662 for 48 h. DAPI‐stained nuclei (blue) and mitochondria marked by TOM20 (green) are shown; scale bar 20 μm. (B, C) Quantification of features of the mitochondrial network: the mean length of summed branches (top panel) and the mean number of network branches (bottom panel) in U2OS (B) and HEK293 (C) Polɣ^D1135A cells. n = 3. (D) Mitochondrial occupancy was measured as the fraction of U2OS Polɣ^D1135A cell area occupied by mitochondrial (TOM20) signal in the experiment shown in A. The average of the untreated and uninduced cells was set to 1; n = 20. (E) The subcellular distribution of mitochondria in the U2OS Polɣ^D1135A cells from the experiment in A was determined as described in the Materials and Methods. The perinuclear and cytosolic zones were defined as reaching 0–2 μm and 2–5 μm, respectively, from the nuclear boundary. The distal zone was defined as any area beyond the distal limit of the cytosolic region. The distribution of mitochondria between these different zones was compared by 2‐way ANOVA; n ≥ 30; error bars in the graphs represent standard deviation. (n, number of biological replicates; ns, not significant; *P ≤ 0.05; **P ≤ 0.01; ****P ≤ 0.0001).