Fig. 7: Mitochondrial actin precedes the recruitment of mitochondrial Dynamins at fusion and fission sites.

a–d Temporal relationship between mitochondrial actin and the fusion dynamin MFN2. Cells were transfected with mCherry-MFN2 and AC-mito while mitochondria were visualised with Mitotracker Deep red. (a) Presence of AC-mito and mCherry-MFN2 at sites of mitochondrial fission, tip-to-side fusion and tip-to-tip fusion. b–d Kinetics of recruitment of AC-mito and mCherry-MFN2 at fission (b) and fusion (c) sites showing recruitment of either marker relative to the time of fusion, as well as mCherry-MFN2 recruitment relative to AC-mito (d). Each point represents an individual event in 4 cells within 3 independent experiments). Bars show the average ± SD. two-sided t-test. e–h Temporal relationship between the fusion dynamin MFN2 and the fission Dynamin DRP1. Cells were transfected with mCherry-MFN2 and GFP-DRP1 while mitochondria were visualised with Mitotracker Deep red. e Presence of GFP-DRP1 and mCherry-MFN2 at sites of mitochondrial fission, tip-to-side fusion and tip-to-tip fusion. (f–h) Kinetics of recruitment of mCherry-MFN2 andGFP-DRP1 at fission (f) and fusion (g) sites showing recruitment of either marker relative to the time of fusion, as well as GFP-DRP1 recruitment relative to mCherry-MFN2 (d). Each point represents an individual event in 3 cells within 3 independent experiments). Bars show the average ± SD. two-sided t-test. (i) Schematic representation of the timing of MFN2 and DRP1 recruitment to tip-to-side fusion (Top) and fission (Bottom) relative to actin recruitment (AC-mito-positive).