The Mic27 subcomplex is specifically dependent on ERMES for assembly but can be restored by ERMES bypass. (A) Representative single planes of deconvolved fluorescence microscopy images of Δmmm1 cells expressing either Mic60-EGFP or Mic27-EGFP (green) and Atp2-mCherry (magenta). (B) Images of Δmic19 mmm1-1 cells coexpressing Mic60-EGFP or Mic27-EGFP (green) and Tim23-mCherry (magenta) and grown at the nonpermissive temperature for the indicated time. (C) Graph depicting the percentage of cells from B with Mic60-EGFP (left) or Mic27-EGFP (right) observed in the indicated appearance relative to Tim23. Data displayed are the summation of three independent experiments with ≥50 cells counted per strain per experiment. (D) Images are shown of Δmic19 mmm1-1 cells (top) or Δmic10 mmm1-1 cells (bottom) coexpressing Atp2-mCherry (magenta) and Mic60-EGFP (green) and grown at the nonpermissive temperature for the indicated time. Arrows mark sites of Mic60 assembly. (E) Serial dilutions of the indicated yeast cells plated on medium containing dextrose (YPD, left) or the nonfermentable carbon source ethanol/glycerol (YPEG, right). (F) Representative images of the indicated strains expressing Mic27-EGFP from E. Arrows mark sites of Mic27 assembly. (G) Graph depicting the percentage of the indicated cells, as shown in Fig. 2 F, with the indicated number of Mic27-EGFP foci per cell. Data represent the average of three independent experiments, and ≥57 cells were counted per strain per experiment. Error bars indicate SEM. Scale bars = 2 µm.