Figure 7.
The mitochondrial fission and fusion machinery perform competing roles in MDC formation. (A) Super-resolution confocal fluorescence microscopy images of a haploid yeast strain expressing sfGFP-Dnm1 from the NOP1 promoter and Tom70-mCherry treated with either DMSO or 200 nM Rap. MDCs are indicated by white arrows. Scale bar = 1 µm. (B) Super-resolution confocal fluorescence microscopy images of a haploid yeast strain expressing sfGFP-Fzo1 from the NOP1 promoter and Tom70-mCherry treated with either DMSO or 200 nM Rap. MDCs are indicated by white arrows. Scale bar = 1 µm. (C) Quantification of the frequency sfGFP-Dnm1 foci or sfGFP-Fzo1 were co-localized with or closely associated with Tom70-mCherry–marked MDCs. Error bars show mean ± SE of three replicates, n ≥ 100 cells per replicate. (D) Quantification of MDC formation in the indicated yeast strains upon treatment with either DMSO, ConcA, or Rap. Error bars show mean ± SE of three replicates, n ≥ 100 cells per replicate. (E) Quantification of the MDC morphologies observed in the indicated yeast strains after Rap treatment shown as a percent of total MDCs. Error bars show mean ± SE of three replicates, n ≥100 cells per replicate. (F and G) Super-resolution confocal fluorescence microscopy images of wild-type (F) or fzo1∆ (G) yeast cells expressing fzo1-1, Tom70-yEGFP, and Tim50-mCherry treated with 200 nM Rap at the indicated temperatures. MDCs are indicated by white arrows. Yellow arrows in G are pointing to Tom70-yEGFP puncta detached from the mitochondrial tubule. Scale bar = 1 µm. (H) Quantification of MDC formation in the indicated yeast strains upon treatment with either DMSO or Rap at the indicated temperatures. Error bars show mean ± SE of three replicates, n ≥ 100 cells per replicate. (I) Model of MDC biogenesis from an OMM extension that forms a double-membrane compartment, elongating and invaginating to form a multilamellar MDC.

The mitochondrial fission and fusion machinery perform competing roles in MDC formation. (A) Super-resolution confocal fluorescence microscopy images of a haploid yeast strain expressing sfGFP-Dnm1 from the NOP1 promoter and Tom70-mCherry treated with either DMSO or 200 nM Rap. MDCs are indicated by white arrows. Scale bar = 1 µm. (B) Super-resolution confocal fluorescence microscopy images of a haploid yeast strain expressing sfGFP-Fzo1 from the NOP1 promoter and Tom70-mCherry treated with either DMSO or 200 nM Rap. MDCs are indicated by white arrows. Scale bar = 1 µm. (C) Quantification of the frequency sfGFP-Dnm1 foci or sfGFP-Fzo1 were co-localized with or closely associated with Tom70-mCherry–marked MDCs. Error bars show mean ± SE of three replicates, n ≥ 100 cells per replicate. (D) Quantification of MDC formation in the indicated yeast strains upon treatment with either DMSO, ConcA, or Rap. Error bars show mean ± SE of three replicates, n ≥ 100 cells per replicate. (E) Quantification of the MDC morphologies observed in the indicated yeast strains after Rap treatment shown as a percent of total MDCs. Error bars show mean ± SE of three replicates, n ≥100 cells per replicate. (F and G) Super-resolution confocal fluorescence microscopy images of wild-type (F) or fzo1∆ (G) yeast cells expressing fzo1-1, Tom70-yEGFP, and Tim50-mCherry treated with 200 nM Rap at the indicated temperatures. MDCs are indicated by white arrows. Yellow arrows in G are pointing to Tom70-yEGFP puncta detached from the mitochondrial tubule. Scale bar = 1 µm. (H) Quantification of MDC formation in the indicated yeast strains upon treatment with either DMSO or Rap at the indicated temperatures. Error bars show mean ± SE of three replicates, n ≥ 100 cells per replicate. (I) Model of MDC biogenesis from an OMM extension that forms a double-membrane compartment, elongating and invaginating to form a multilamellar MDC.

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