Figure 1.

Characterization of mitochondrial fission-related protein KO HeLa cell lines. (A) Cytosol and mitochondrial fractions were analyzed by Western blotting. Asterisks, nonspecific bands. (B) Confocal images of the indicated cell lines; Drp1 (green) and Tom20 (red). Bar, 20 µm. Insets, magnified images of the boxed regions. Bar, 5 µm. Bottom right, fluorescence obtained for the Drp1 channel. Heat map, Drp1 fluorescence intensity (FI). (C) Western blotting of mitochondrial fractions for Drp1 and mitofilin. Histogram, quantification of three independent biological replicates. n = 3. Data represent mean ± SD. (D) Drp1 fluorescence per focus on the MOM. 200 foci were analyzed from randomly selected regions for each group in B. n = 200. In C and D, the data were normalized to the wild-type cells. Data represent mean ± SD. (E) Confocal images of the indicated cells; cytochrome c (green) and PMP70 (red; a peroxisomal marker). Bar, 10 µm. (F) Subcellular localization of MiD51-GFP. Mff-KO cells were transfected with MiD51-GFP and immunostained with the indicated antibodies. Bars, 10 µm. Magnified image of the boxed region is shown as an inset. Bar, 2.5 µm. Wt, wild type.