Fig. 2

Mitochondrial fission reduces membrane order. a, b Fluorescence images of SUM159- (a) or MDA-MB-231- (b) WT and PISD cells stably expressing mitochondrially targeted GFP (Mito-GFP) treated with vehicle control, α-lysophosphatidylethanolamine (LPE, 50 μM), or leflunomide (Lef, 50 μM) for 48 h. Scale bar = 10 μm. c, d Graphs show the mean fraction + SD of fissioned or fused mitochondria after treatment with vehicle control, LPE, or Lef (n ≥ 13 images for each treatment group) in SUM159 (c) or MDA-MB-231 (d) cells. *p < 0.05. **p < 0.01. ***p < 0.0001. e Representative images of SUM159-WT, WT treated with α-lysophosphatidylethanolamine (LPE, 50 μM), PISD, PISD treated with leflunomide (Lef, 50 μM), or Drp1 cells stained with laurdan. Images show pseudocolored general polarization (GP) values. Red color indicates areas of high membrane order and less dynamics, whereas blue color denotes areas of low membrane order and more dynamics. Gray color indicates areas that fell between the ordered and disordered phases based upon an arbitrary cutoff used for all groups. Arrows point to cell membranes. Scale bar = 50 μm. f Graph shows mean + SD for the percentage of all ordered and disordered pixels for each image in e (n = 10 images per group). *p < 0.05. **p < 0.01. ***p < 0.0001