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. 2016 Jun 3;17(7):1029–1043. doi: 10.15252/embr.201541832

Figure 4. A role for septins in mitochondrial fission.

Figure 4

  1. HeLa cells expressing SEPT6‐GFP were stained with MitoTracker Red CMXRos and fixed for confocal microscopy. Inset image highlights the location of septins at the sites of mitochondrial fission. The scale bar represents 5 μm.
  2. HeLa cells were fixed and labelled with antibody against SEPT7 and secondary antibody coupled to HRP, and processed for TEM. The right image is enlarged from the boxed region in the TEM image and shows a SEPT7 (S7) filament (labelled in black) intersecting with the mitochondrial (Mt) fission site. The scale bar represents 100 nm. See also Fig EV4.
  3. HeLa cells were stained with MitoTracker Red CMXRos for live SOFI‐based super‐resolution microscopy. Each image was reconstructed from 100 raw TIRF frames using a custom‐made SOFI‐based algorithm. Arrows highlight SEPT6 association with the location of mitochondrial fission. The scale bar represents 1 μm. See also Movie EV4.
  4. HeLa cells were labelled with MitoTracker Red CMXRos, fixed for widefield microscopy and labelled with antibodies to SEPT7 and Drp1. Arrows highlight one example of Drp1 and SEPT7 association with the location of mitochondrial fission. The scale bar represents 1 μm.
  5. HeLa cells were transfected with Mito‐BFP and ActA‐SEPT7 mCherry, fixed and labelled with antibody to Drp1 for widefield microscopy. The scale bar represents 5 μm. Inset images highlight Drp1 and SEPT7 associated with the sites of mitochondrial fission. Pearson's correlation coefficient from three independent experiments (mean ± SEM) shows that the mitochondria in ActA‐SEPT7‐transfected cells recruit significantly more Drp1 than control cells (Mito‐BFP transfected alone). Student's t‐test, *P < 0.05.