Figure S3.
CCDC51 overexpression leads to mitochondrial fragmentation and its accumulation at sites that do not co-localize with autophagy markers. (A) Representative single-plane confocal images of U2OS cells transiently transfected with untagged CCDC51, fixed, and immunolabeled for HSP60 and CCDC51. The white asterisk marks a non-transfected cell with tubular mitochondrial morphology and endogenous distribution of CCDC51. The yellow asterisk marks a CCDC51-overexpressing cell with fragmented mitochondria in which CCDC51 concentrates at discrete sites (examples marked with yellow arrows). Numbered insets at right correspond to the dashed boxes shown at left. (B) Representative maximum projection confocal images of HeLa wild-type and Drp1 KO cells expressing empty vector (top) or overexpressing GFP-CCDC51 (bottom) that were fixed and immunolabeled for TOMM20. Images shown correspond to quantification shown in Fig. 3 G. Arrows mark examples of GFP-CCDC51 focal enrichment. (C) Representative maximum intensity projection confocal images are shown of wild-type HeLa cells expressing empty vector (top) or GFP-CCDC51 (bottom) and co-expressing TIMM50-mCherry and BFP-OMP25. Arrows mark examples of focal accumulation of GFP-CCDC51. See Fig. 3 H for corresponding images in HeLa Drp1 KO cells. (D) Representative maximum intensity projection confocal images are shown of a U2OS cell overexpressing GFP-CCDC51 and mCherry-LC3 that was fixed and immunolabeled for HSP60. Insets at right correspond to dashed boxes shown at left. Note that GFP-CCDC51 foci do not appear to associate with mCherry-LC3. Scale bars: (A and D) 15 µm (3 µm in insets); (B and C) 5 µm.

CCDC51 overexpression leads to mitochondrial fragmentation and its accumulation at sites that do not co-localize with autophagy markers. (A) Representative single-plane confocal images of U2OS cells transiently transfected with untagged CCDC51, fixed, and immunolabeled for HSP60 and CCDC51. The white asterisk marks a non-transfected cell with tubular mitochondrial morphology and endogenous distribution of CCDC51. The yellow asterisk marks a CCDC51-overexpressing cell with fragmented mitochondria in which CCDC51 concentrates at discrete sites (examples marked with yellow arrows). Numbered insets at right correspond to the dashed boxes shown at left. (B) Representative maximum projection confocal images of HeLa wild-type and Drp1 KO cells expressing empty vector (top) or overexpressing GFP-CCDC51 (bottom) that were fixed and immunolabeled for TOMM20. Images shown correspond to quantification shown in Fig. 3 G. Arrows mark examples of GFP-CCDC51 focal enrichment. (C) Representative maximum intensity projection confocal images are shown of wild-type HeLa cells expressing empty vector (top) or GFP-CCDC51 (bottom) and co-expressing TIMM50-mCherry and BFP-OMP25. Arrows mark examples of focal accumulation of GFP-CCDC51. See Fig. 3 H for corresponding images in HeLa Drp1 KO cells. (D) Representative maximum intensity projection confocal images are shown of a U2OS cell overexpressing GFP-CCDC51 and mCherry-LC3 that was fixed and immunolabeled for HSP60. Insets at right correspond to dashed boxes shown at left. Note that GFP-CCDC51 foci do not appear to associate with mCherry-LC3. Scale bars: (A and D) 15 µm (3 µm in insets); (B and C) 5 µm.

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