Figure S2.
MDC formation requires ERMES and Gem1, but not other organelle contact site proteins (related to Fig. 3).(A) Quantification of rapamycin-induced MDC formation in WT and gem1Δ cells. Oac1-positive, Tim50-negative MDCs were quantified. Error bars show mean ± SEM of three replicates, n = 50–100 cells per replicate. (B) Quantification of concanamycin A–induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (C) Quantification of cycloheximide-induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (D) Quantification of rapamycin-induced MDC formation in WT and lam6Δ cells. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (E) Quantification of Rap-induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (F) Indirect immunofluorescence of cells expressing EV or overexpressing GEM1 or the indicated GEM1 mutant and Mdm34-GFP or Tom70-GFP. Top row images (Gem1) were acquired with the same settings and are nonadjusted. Asterisk (Gem1*) indicates that the fluorescence intensity was increased. Images show single focal planes. Merged images show Gem1* and Mdm34-GFP or Tom70-GFP. Scale bars = 2 µm. CHX, cycloheximide; ConcA, concanamycin A; Rap, rapamycin.

MDC formation requires ERMES and Gem1, but not other organelle contact site proteins (related to Fig. 3 ). (A) Quantification of rapamycin-induced MDC formation in WT and gem1Δ cells. Oac1-positive, Tim50-negative MDCs were quantified. Error bars show mean ± SEM of three replicates, n = 50–100 cells per replicate. (B) Quantification of concanamycin A–induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (C) Quantification of cycloheximide-induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (D) Quantification of rapamycin-induced MDC formation in WT and lam6Δ cells. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (E) Quantification of Rap-induced MDC formation in WT and the indicated mutant yeast. Error bars show mean ± SEM of three replicates, n = 100 cells per replicate. (F) Indirect immunofluorescence of cells expressing EV or overexpressing GEM1 or the indicated GEM1 mutant and Mdm34-GFP or Tom70-GFP. Top row images (Gem1) were acquired with the same settings and are nonadjusted. Asterisk (Gem1*) indicates that the fluorescence intensity was increased. Images show single focal planes. Merged images show Gem1* and Mdm34-GFP or Tom70-GFP. Scale bars = 2 µm. CHX, cycloheximide; ConcA, concanamycin A; Rap, rapamycin.

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