Figure EV1. Bioenergetics defects in Mfn2 KO cells are independent of mitochondrial morphology.

• A
  ATP levels in the indicated cell lines when metabolic flexibility is allowed (n = 3 independent experiments). Data are presented as mean ± SEM.
• B
  Representative Western blots of the indicated proteins in Mfn2 KO fibroblasts transfected with plasmids expressing Mfn1, DN‐Drp1, or a control plasmid (CT) (n = 3 independent experiments).
• C–F
  Expression of Mfn1 restores mitochondrial morphology in Mfn2 KO MEFs. WT or Mfn2 KO MEFs were co‐transfected with plasmids encoding mt‐RFP and Mfn1 or control plasmids. (C) Representative images. Scale bar = 20 µm. (D) Percentage of cells displaying globular mitochondria (n = 54–414 cells analyzed in three independent experiments), (E) aspect ratio, and (F) circularity (n = 260–300 mitochondria analyzed in three independent experiments). Data are presented as mean ± SEM.
• G–J
  Expression of DN‐Drp1 restores mitochondrial morphology in Mfn2 KO MEFs. WT or Mfn2 KO MEFs were co‐transfected with plasmids encoding mt‐RFP and DN‐Drp1 or control plasmids. (G) Representative images. Scale bar = 20 µm. (H) Percentage of cells displaying globular mitochondria (n = 135–470 cells analyzed in three independent experiments), (I) aspect ratio and (J) circularity (n = 300 mitochondria were analyzed in three independent experiments). Data are presented as mean ± SEM.
• K, L
  Restoration of mitochondrial morphology by expression of (K) Mfn1 or (L) DN‐Drp1 in Mfn2 KO MEFs has no effect on mitochondrial membrane potential (n = 90 cells analyzed in three independent experiments). Data are presented as mean ± SEM.
• M, N
  Restoration of mitochondrial morphology by expression of (M) Mfn1 or (N) DN‐Drp1 in Mfn2 KO MEFs has no effect on basal oxygen consumption rate (n = 3 independent experiments). Data are presented as mean ± SEM.

Data information: *P < 0.05, one‐way ANOVA followed by Tukey’s post hoc test.