Prolonged CCDC51 depletion leads to mitochondrial hyperfusion that precedes formation of lamellar mitochondria. (A) Single-plane deconvolved images are shown of U2OS cells at the indicated times after treatment with the indicated siRNA oligonucleotides and stained with MitoTracker Red. Insets correspond to dashed boxes. Images correspond to quantification shown in Fig. 2 E. (B) Western blot analysis with the indicated antibodies of lysate from cells treated as in A and Fig. 2 E. (C) Western blot analysis with the indicated antibodies of lysate from cells corresponding to Fig. 2, F and G. (D) Single-plane deconvolved images are shown of HeLa cells at the indicated times after treatment with the indicated siRNA oligonucleotides and stained with MitoTracker Red. Insets correspond to dashed boxes. Yellow arrows mark mitochondrial bulbs and white arrows mark lamellar mitochondria. (E) Western blot analysis with the indicated antibodies of whole-cell lysate from cells treated as in D. (F) A graph is shown of mitochondrial morphology characterization from cells as in D and E. Data shown represents 50–100 cells per condition in each of three independent experiments, and bars indicate SEM. The P value shown represents an unpaired two-tailed t test of lamellar mitochondrial morphology. (G) A graph is shown of mitochondrial morphology characterization of cells treated with the indicated siRNA for 120 h. Data represent an average of results from two independent experimental replicates with at least 75 cells counted per experiment. Scale bars: (A) 15 µm (3 µm in inset); (D) 15 µm (5 µm in inset). Source data are available for this figure: SourceData FS2.