Figure 1.
Mdm33 and CCDC51 are conserved mediators of mitochondrial morphology. (A) A schematic depicting the predicted domain architectures of yeast Mdm33 and human CCDC51. (B) Western analysis with the indicated antibodies of whole-cell lysates from U2OS CRISPRi cells expressing control sgRNA or sgRNAs targeting CCDC51. (C) Representative maximum intensity projection confocal images of U2OS CRISPRi cells expressing control or CCDC51-targeted sgRNAs and stained with MitoTracker Red. Insets below are single-plane images and correspond to the indicated dashed boxes. Arrows mark discontinuities in MitoTracker staining. (D) A graph of mitochondrial morphology categorization from the indicated CRISPRi cells as in C. Data shown represent 100 cells per condition in each of three independent experiments, and bars indicate SEM. Asterisks (****P < 0.0001) represent unpaired two-tailed t tests of tubular mitochondrial morphology. (E) Representative maximum intensity projection confocal images are shown of the indicated CRISPRi cells expressing mCherry-OMP25, TIMM50-GFP, and mito-HaloTag labeled with JF646. (F) Maximum intensity projection confocal images are shown of the indicated yeast strains expressing Tom20-mCherry, Pam17-EGFP, and mito-TagBFP. Dashed lines indicate cell outlines. (G) EM images are shown of mitochondria from U2OS CRISPRi cells expressing control or CCDC51 sgRNA, where indicated. Yellow arrows mark internal ring structures and white arrows mark cristae. (H) Maximum intensity projection confocal images are shown of the indicated yeast strains-expressing Tom20-EGFP and mito-dsRed. Human CCDC51 is expressed under the control of an estradiol-driven promoter, and all strains were grown constitutively in the presence of 20 nM estradiol. Dashed lines indicate cell outlines. (I) A graph of the categorization of mitochondrial morphology from cells as in H. Data shown represent 75 cells per condition in each of three independent experiments, and bars indicate SEM. Asterisks (***P < 0.001) represent an unpaired two-tailed t test of tubular mitochondrial morphology. Scale bars = (C) 15 µm (5 µm in insets); (E and F) 3 µm; (G) 800 nm; and (H) 3 µm. Source data are available for this figure: SourceData F1.

Mdm33 and CCDC51 are conserved mediators of mitochondrial morphology. (A) A schematic depicting the predicted domain architectures of yeast Mdm33 and human CCDC51. (B) Western analysis with the indicated antibodies of whole-cell lysates from U2OS CRISPRi cells expressing control sgRNA or sgRNAs targeting CCDC51. (C) Representative maximum intensity projection confocal images of U2OS CRISPRi cells expressing control or CCDC51-targeted sgRNAs and stained with MitoTracker Red. Insets below are single-plane images and correspond to the indicated dashed boxes. Arrows mark discontinuities in MitoTracker staining. (D) A graph of mitochondrial morphology categorization from the indicated CRISPRi cells as in C. Data shown represent 100 cells per condition in each of three independent experiments, and bars indicate SEM. Asterisks (****P < 0.0001) represent unpaired two-tailed t tests of tubular mitochondrial morphology. (E) Representative maximum intensity projection confocal images are shown of the indicated CRISPRi cells expressing mCherry-OMP25, TIMM50-GFP, and mito-HaloTag labeled with JF646. (F) Maximum intensity projection confocal images are shown of the indicated yeast strains expressing Tom20-mCherry, Pam17-EGFP, and mito-TagBFP. Dashed lines indicate cell outlines. (G) EM images are shown of mitochondria from U2OS CRISPRi cells expressing control or CCDC51 sgRNA, where indicated. Yellow arrows mark internal ring structures and white arrows mark cristae. (H) Maximum intensity projection confocal images are shown of the indicated yeast strains-expressing Tom20-EGFP and mito-dsRed. Human CCDC51 is expressed under the control of an estradiol-driven promoter, and all strains were grown constitutively in the presence of 20 nM estradiol. Dashed lines indicate cell outlines. (I) A graph of the categorization of mitochondrial morphology from cells as in H. Data shown represent 75 cells per condition in each of three independent experiments, and bars indicate SEM. Asterisks (***P < 0.001) represent an unpaired two-tailed t test of tubular mitochondrial morphology. Scale bars = (C) 15 µm (5 µm in insets); (E and F) 3 µm; (G) 800 nm; and (H) 3 µm. Source data are available for this figure: SourceData F1.

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