Extended Data Figure 2. Adenovirally-expressed Gly-substituted mini-peptides derived from Mfn2 HR1 specifically regulate Mfn-mediated mitochondrial fusion.

a. Changes in mitochondrial aspect ratio provoked by adeno-367-384Gly and adeno-398-418Gly in MEFs having different Mfn expression profiles, compared to adeno-WT Mfn2 and adeno-(Mfn2) HR1. Each point is the aspect ratio of an individual mitochondrion; Group data are meanαSEM; *=P<0.05 vs adeno-null (ANOVA). Exact numbers of mitochondria measured from 4 or 5 separate experiments per condition are shown on the figure. b. Mitochondrial polarization status and aspect ratio in WT and Mfn2 KO MEFs expressing WT Mfn2 or fragments thereof. Each point is the mean of ~20 mitochondria from ~5 cells in the number of independent experiments indicated on the graph. Group data are meanαSEM; *=P<0.05 vs ad-null Ctrl (ANOVA). Representative (of 15–40 images/group) merged confocal images of WT MEFs infected with ad-Mfn2 or the two biologically active Ad-peptides are to the right; scale bar is 10 microns. c. MitoTracker Green and m-cherry Parkin (top) or Lyso-Red (bottom) co-stained WT or Mfn2 KO MEFs before (left) or 1 hour after (right) treatment with 50 nM FCCP. Group data are meanαSEM; there were no differences between groups (ANOVA; n=3 independent experiments per condition). Scale bars are 10 µm.