Injury-triggered mitochondrial fragmentation is conserved in HeLa cells. (A–C) Images showing mitochondrial shape in response to PM injury in WT (A), EGTA-treated WT (B), and DRP1 KO (C) HeLa cells. In all cases, mitochondria are marked by TOM20, a structural protein of the OMM. Arrows mark the site of injury. Boxes indicate the area of zoom shown below. Scale bar = 10 µm (whole cell) and 5 µm (zoom). (D) Plot showing FM 1-43 dye intensity after injury in WT (n = 31) and DRP1 KO (n = 44) HeLa cells. (E) Plot showing percentage of cells that failed to repair in independent experiments (n = 3). (F) Plot showing Lifeact-mCherry intensity after PM injury in WT (n = 5) and DRP1 KO (n = 7) HeLa cells. *, P < 0.05 by unpaired t test. Error bars represent SEM.