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Figure 3


Fig. 3. Mitochondrial nucleoids undergo remodeling. (A-C) Quantification of micrococcal-nuclease-sensitivity assays of (A) 21S_rRNA, (B) ATP6 and (C) intergenic region 81 using ~1 mg of toluene-permeabilized mitochondria isolated from BY4741 cells grown in glucose-containing (bullet) and glycerol-containing ({circ}) medium. Representative autoradiograms are shown in supplementary material Fig. S4. (D) Western blot of Pgk1, Abf2 and Cox2 proteins from cell extracts of BY4741 cells grown in YPD (glu) and YPG (gly) medium. Because expression of Pgk1 is not dependent on a carbon source (Roberts and Hudson, 2006), an anti-Pgk1 antibody was used to estimate the protein loading. Unlike Cox2 – the expression of which is much higher in glycerol than in glucose medium – Abf2 levels were comparable in both YPD and YPG. (E) Relative mtDNA copy number in BY4741 cells grown in YPD (glucose) and YPG (glycerol) medium assessed by Southern blot hybridizations using probes derived from the mitochondrial 21S_RNA and nuclear ACT1 genes. Error bars, average deviations.





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