Figure 4.
Genetic suppressors of ERMES mutants differ in their ability to restore MDC formation.(A) Quantification of rapamycin-induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (B) Quantification of Rap-induced MDC formation in WT, mmm1Δ, and mdm12Δ cells expressing EV or overexpressing MMM1, MMM1(L247S), MDM12, or MDM12(V214S). Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (C) Widefield images of WT and the indicated mutant yeast expressing Tom70-GFP and Tim50-mCherry and EV or VPS13(D716H) treated with DMSO or rapamycin. Arrows mark MDCs. Images show maximum intensity projections of merged Tom70-GFP and Tim50-mCherry. Scale bars = 2 µm. (D) Quantification of C. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (E) Widefield images of WT and mmm1Δ yeast expressing Tom70-GFP and Tim50-mCherry and EV or overexpressing MCP1 treated with DMSO or Rap. Arrows mark MDCs. Images show maximum intensity projections of merged Tom70-GFP and Tim50-mCherry. Scale bars = 2 µm. (F) Quantification of E. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (G) Quantification of rapamycin-induced MDC formation in WT and gem1Δ cells expressing EV or overexpressing MCP1. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. See also Fig. S3. Rap, rapamycin.

Genetic suppressors of ERMES mutants differ in their ability to restore MDC formation. (A) Quantification of rapamycin-induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (B) Quantification of Rap-induced MDC formation in WT, mmm1Δ, and mdm12Δ cells expressing EV or overexpressing MMM1, MMM1(L247S), MDM12, or MDM12(V214S). Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (C) Widefield images of WT and the indicated mutant yeast expressing Tom70-GFP and Tim50-mCherry and EV or VPS13(D716H) treated with DMSO or rapamycin. Arrows mark MDCs. Images show maximum intensity projections of merged Tom70-GFP and Tim50-mCherry. Scale bars = 2 µm. (D) Quantification of C. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (E) Widefield images of WT and mmm1Δ yeast expressing Tom70-GFP and Tim50-mCherry and EV or overexpressing MCP1 treated with DMSO or Rap. Arrows mark MDCs. Images show maximum intensity projections of merged Tom70-GFP and Tim50-mCherry. Scale bars = 2 µm. (F) Quantification of E. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (G) Quantification of rapamycin-induced MDC formation in WT and gem1Δ cells expressing EV or overexpressing MCP1. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. See also Fig. S3. Rap, rapamycin.

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