Figure 1.
Rapid and localized mitochondrial fragmentation occurs in response to PM injury. (A) WT MEF showing mitochondria labeled with MitoTracker Green before and after PM injury. Red box indicates the area magnified on the right. (B) Schematic showing mitochondrial fragmentation analysis. Cells were injured by pulsed laser (left); dotted box (right) marks the mitochondria that responded in the injured cell. Extent of fragmentation 30 s after injury was calculated by dividing the area of injured cells with fragmented (green) mitochondria by the area of cells containing the responding mitochondria (dotted box). (C) FM 1-43 dye intensity after injury of untreated (n = 17) or EGTA-treated (n = 21) WT MEFs. (D) Percentage of cells that failed to repair (n = 3 experiments). (E) Images showing mitochondrial shape in response to PM injury in EGTA-treated WT MEFs. (F) Images showing injury-induced change in mitochondrial membrane potential in a mitochondrion labeled with TMRE and TOM20-YFP (Mito). Arrows indicate sites of fragmentation. (G) Heat map images showing change in TMRE fluorescence intensity after injury. (H) Plot showing the ratio of TMRE/TOM20-YFP fluorescence intensity of cell shown in G. (I) Images showing shape of injury-proximal mitochondria during PMR in WT MEFs treated with ruthenium red or mitoTEMPO. (J) Extent of mitochondrial fragmentation at injury site (n = 3 experiments). Unless otherwise noted, arrows mark the site of injury. Scale bar = 10 µm (A, E, and G) or 5 µm (A inset and I) or 1 μm (F). *, P < 0.05 by unpaired t test. Error bars represent SEM.

Rapid and localized mitochondrial fragmentation occurs in response to PM injury. (A) WT MEF showing mitochondria labeled with MitoTracker Green before and after PM injury. Red box indicates the area magnified on the right. (B) Schematic showing mitochondrial fragmentation analysis. Cells were injured by pulsed laser (left); dotted box (right) marks the mitochondria that responded in the injured cell. Extent of fragmentation 30 s after injury was calculated by dividing the area of injured cells with fragmented (green) mitochondria by the area of cells containing the responding mitochondria (dotted box). (C) FM 1-43 dye intensity after injury of untreated (n = 17) or EGTA-treated (n = 21) WT MEFs. (D) Percentage of cells that failed to repair (n = 3 experiments). (E) Images showing mitochondrial shape in response to PM injury in EGTA-treated WT MEFs. (F) Images showing injury-induced change in mitochondrial membrane potential in a mitochondrion labeled with TMRE and TOM20-YFP (Mito). Arrows indicate sites of fragmentation. (G) Heat map images showing change in TMRE fluorescence intensity after injury. (H) Plot showing the ratio of TMRE/TOM20-YFP fluorescence intensity of cell shown in G. (I) Images showing shape of injury-proximal mitochondria during PMR in WT MEFs treated with ruthenium red or mitoTEMPO. (J) Extent of mitochondrial fragmentation at injury site (n = 3 experiments). Unless otherwise noted, arrows mark the site of injury. Scale bar = 10 µm (A, E, and G) or 5 µm (A inset and I) or 1 μm (F). *, P < 0.05 by unpaired t test. Error bars represent SEM.

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