Fig. 4. Ncr1p deficiency decreases mitochondrial function and dynamics.
S. cerevisiae BY4741 and ncr1Δ::KanMX4 cells were grown in SC-glucose medium.
A. Oxygen consumption rates were measured in exponential (log) and PDS phase cells. Values are mean ± SD of at least three independent experiments. ****p<0.0001, *p<0.05; Two-way ANOVA and Bonferroni test.
B. Cytochrome c oxidase (COX) specific-activity was measured in cells grown to PDS phase. Values are mean ± SD of at least three independent experiments. ****p<0.0001, unpaired Student’s t-test.
C. Immunoblot analysis of mitochondrial porin levels in cells grown to PDS phase. For each lane the Por1p signal was normalized to the signal for the Pgk1p internal standard (value shown below each lane). A representative experiment out of three is shown.
D. Cells were grown to exponential phase and fivefold serial dilutions were plated in solid medium containing glucose or glycerol as carbon source. One representative experiment out of three is shown.
E. Mitochondrial membrane potential was determined by flow cytometry using cells grown to PDS phase, unlabeled (auto fluorescence) or labeled with DiOC6(3). Representative histograms for each condition are shown in Supplemental Fig. S3. Values are mean ± SD of three independent experiments. ***p<0.001, unpaired Student’s t-test.
F. S. cerevisiae BY4741 and ncr1Δ::KanMX4 cells transformed with pYX222-mtDsRed (expressing mitochondrial DsRed) were grown to exponential (log) and PDS phase. Live cells were visualized by fluorescence microscopy. One representative experiment out of three is shown. Scale bar: 5 μm.