Figure 6.

CoFe granule number varies with quality of carbon source

(A) z-stacked images of PDC1-MS2 or ENO1-MS2 mRNA in strains coexpressing the MS2-CP-GFP fusion, grown in SC media with either 2% glucose, 2% raffinose, 2% galactose, or 3% ethanol. Scale bar: 2 μm.

(B) Quantification of mRNA granules per cell for PDC1 and ENO1 mRNA in strains grown in the different carbon sources (Gluc = Glucose, Raff = Raffinose, Gal = Galactose, EtOH = Ethanol). n = 50. Error bars are ±SD. ∗p < 0.005 relative to other columns from a 1-way ANOVA and the Tukey's HSD test.

(C) Graph representing the gene expression ratio of PDC1 or ENO1 in strains (yMK1586 and yMK2468) grown in the different carbon sources. The gene expression ratio was calculated according to the Pfaffl method. This approach considers the PCR efficiency for the different genes and is expressed as the change in target gene levels (PDC1 and EN O1) between glucose conditions and the different carbon sources over the change in reference gene levels (ACT1) between glucose conditions and the different carbon sources (Pfaffl, 2001) (Gluc = Glucose, Raff = Raffinose, Gal = Galactose, EtOH = Ethanol). Error bars are ±SD.

(D) z-stacked images of FBA1, GPM1, and TPI1 mRNAs probed using smFISH in DAPI-stained wild-type strains. Strains were grown in SC media with either 2% glucose or 3% ethanol. Scale bar: 1 μm.

(E) Graph representing the number of mRNA molecules within each smFISH foci in cells grown in either glucose (black) or ethanol (EtOH; magenta). Foci were separated into either all foci or multi mRNA foci dependent upon the number of mRNAs predicted to reside within that foci (see Methods). Box and line represent the interquartile range and the median, respectively

(F) Quantification of total mRNA molecules per cell in cells grown in either glucose (black) or ethanol (EtOH; magenta), as measured by smFISH. Box and line represent the interquartile range and the median, respectively

(G) Quantification of the number of multi-mRNA containing foci per cell in cells grown in either glucose (black) or ethanol (EtOH; magenta). Box and line represent the interquartile range and the median, respectively