Decreased AdoMet levels result in increased SG formation. (A) Percentage of cells forming foci in strains endogenously expressing either mutant or WT Sam1-GFP with RFP-tagged metabolic enzymes (Sam2, Ade17) and SG markers (Ded1, Pbp1) in 1-d cultures. (B) Western blot analysis from strains used in A at 1-d time point. Numbers below each mutant indicate clone number for that genotype. (C) Quantification of proteins levels from B. Protein expression for each protein is normalized to the WT sample. (D) Quantification of mRNA levels using qPCR analysis from WT and mutant Sam1 alleles. (E) Percentage of cells forming foci in strains supplemented with 250 µM AdoMet and endogenously expressed WT or mutant Sam1-GFP alleles and Ded1-mCherry at 1-d growth. (F) Western blot analysis from strains used in E at 1-d time point. (G) Quantification of proteins levels from E. Protein expression is normalized to non–AdoMet-treated Sam1 and Ded1 within their respective genotype. Bar graphs in A, C–E, and G are presented as average ± SEM of three independent replicates. Pgk1 was used as a loading control in B and F. Statistical significance in A, C–E, and G was determined by one-tailed paired Student’s t test (*, P < 0.05).