Figure 3.
Dnm1 fails to complete mitochondrial division in the absence of Mdi1. (A) Maximum intensity projections of deconvolved images of the indicated yeast strains expressing mito-dsRed grown to log phase and either directly imaged (top) or treated for 40 min with 25 µM CCCP (bottom) prior to imaging. Arrows mark sites of mitochondrial constriction. (B) Maximum intensity projections of deconvolved images of the indicated yeast strains expressing chromosomally integrated EGFP-Fis1 (green) and mito-DsRed (magenta). Arrows mark sites of focal accumulation of Fis1. (C) As in B for cells expressing an endogenous chromosomal Dnm1-EGFP tag (green). Arrows mark sites of Dnm1 association with mitochondria. (D) As in C for cells treated for 30 min with 25 µM CCCP prior to imaging. Dashed yellow line corresponds to the fluorescence intensity linescan shown below. (E) Single plane (left) and maximum intensity projection (right) confocal fluorescence microscopy images are shown of the indicated yeast strains expressing Tom20-mCherry (OMM, magenta), mito-TagBFP (matrix, blue), and Dnm1-EGFP (green) and treated for 40 min with 25 µM CCCP prior to imaging. Arrows mark sites of Dnm1-marked hyperconstriction where the OMM label appears continuous. Dashed yellow lines correspond to the fluorescence intensity linescans shown at right. (F) Maximum intensity projections of confocal microscopy images of the indicated yeast strains expressing chromosomally tagged Tom20-mCherry (OMM, magenta) and Pam17-EGFP (IMM, green). (G) As in F for the indicated yeast strains treated for 45 min with 25 µM CCCP. Dashed yellow lines correspond to the fluorescence intensity linescans shown at right. Quantification shown in the figure represents the percentage of Tom20-positive mitochondrial hyperconstrictions that appeared positive for Pam17-EGFP. Cell boundaries are indicated with dotted lines. Scale bars = 2 µm (E), 3 µm (A–D and F–G).

Dnm1 fails to complete mitochondrial division in the absence of Mdi1. (A) Maximum intensity projections of deconvolved images of the indicated yeast strains expressing mito-dsRed grown to log phase and either directly imaged (top) or treated for 40 min with 25 µM CCCP (bottom) prior to imaging. Arrows mark sites of mitochondrial constriction. (B) Maximum intensity projections of deconvolved images of the indicated yeast strains expressing chromosomally integrated EGFP-Fis1 (green) and mito-DsRed (magenta). Arrows mark sites of focal accumulation of Fis1. (C) As in B for cells expressing an endogenous chromosomal Dnm1-EGFP tag (green). Arrows mark sites of Dnm1 association with mitochondria. (D) As in C for cells treated for 30 min with 25 µM CCCP prior to imaging. Dashed yellow line corresponds to the fluorescence intensity linescan shown below. (E) Single plane (left) and maximum intensity projection (right) confocal fluorescence microscopy images are shown of the indicated yeast strains expressing Tom20-mCherry (OMM, magenta), mito-TagBFP (matrix, blue), and Dnm1-EGFP (green) and treated for 40 min with 25 µM CCCP prior to imaging. Arrows mark sites of Dnm1-marked hyperconstriction where the OMM label appears continuous. Dashed yellow lines correspond to the fluorescence intensity linescans shown at right. (F) Maximum intensity projections of confocal microscopy images of the indicated yeast strains expressing chromosomally tagged Tom20-mCherry (OMM, magenta) and Pam17-EGFP (IMM, green). (G) As in F for the indicated yeast strains treated for 45 min with 25 µM CCCP. Dashed yellow lines correspond to the fluorescence intensity linescans shown at right. Quantification shown in the figure represents the percentage of Tom20-positive mitochondrial hyperconstrictions that appeared positive for Pam17-EGFP. Cell boundaries are indicated with dotted lines. Scale bars = 2 µm (E), 3 µm (A–D and F–G).

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