Mic60 assemblies specifically redistribute in proximity to induced vCLAMPs. (A) Images of Δmic19 cells constitutively overexpressing mKate-Vps39 (magenta) and expressing mito-TagBFP (blue) and Mic60-EGFP (green). White arrows mark Mic60-EGFP enrichment in proximity to the vCLAMP. (B) Images as in A for Δmic60 cells expressing Mic27-EGFP (green). Yellow arrows mark Mic27 assemblies not associated with a vCLAMP. (C) Graph of Mic60 and Mic27 assemblies that colocalize or localize adjacent to vCLAMPs from cells as in A and B. Data are the summation of the indicated number of MICOS assemblies from three independent experiments. (D) Images of otherwise wild-type cells constitutively overexpressing mKate-Vps39 (magenta) and expressing mito-TagBFP (blue) and Mic60-EGFP (green). White arrows mark Mic60 enrichment relative to mito-TagBFP in proximity to the vCLAMP. (E) Images as in D for cells expressing Mic27-EGFP (green). Yellow arrows mark vCLAMP sites with no discernable Mic27 enrichment. (F) Graph of vCLAMPs from cells as in D and E where Mic60-EGFP or Mic27-EGFP are discernably enriched compared with mito-TagBFP. Data are the average of three independent experiments, and >35 vCLAMPs were counted per experiment. Error bars indicate SEM. (G) Model for the role of interorganelle MCSs in MICOS subcomplex assembly and positioning. Cells are outlined with dashed lines. Scale bars = 2 µm.