Figure 5.

Set4 promotes activation of stress response genes. A, GO analysis of significantly differentially expressed genes identified by microarray analysis of set4Δ cells from Kemmeren et al. (35). The −log₁₀ p value of the enriched GO biological process categories are shown for both down-regulated and up-regulated genes. B, RT-qPCR of stress response genes identified by Kemmeren et al. (35) from yeast with empty pMN3 or pMN3-SET4 in the presence or absence of β-estradiol. Expression levels were normalized to SCR1. -Fold change relative to empty vector strain without β-estradiol treatment is shown. Error bars, S.E. from at least three biological replicates. C, RT-qPCR of stress response genes from WT (yEG001) and set4Δ (yEG322) strains grown in YPD and treated with 0.4 mm H₂O₂ for 30 min. Expression levels were normalized to SCR1. -Fold change relative to the WT strain without H₂O₂ treatment is shown. Error bars, S.E. from at least three biological replicates. PNC1 consistently showed lower expression in set4Δ cells treated with H₂O₂, although this did not reach the threshold for statistical significance due to variability in its expression levels in H₂O₂. D, -fold change, as in C, of SET4 mRNA in WT cells without or with 0.4 mm H₂O₂ for 30 min. E, chIP of FLAG-Set4 (expressed from its endogenous promoter; yEG513) from cells grown to mid-log phase in YPD or YPD with 0.4 mm H₂O₂ for 30 min. Percentage of input was calculated for each IP and primer set, and enrichment is shown for FLAG-Set4 relative to the WT (untagged) strain either with or without H₂O₂ treatment. The primers detect the promoter sequences for each of the indicated genes. Error bars, S.E. from three biological replicates. For all panels, asterisks represent p values from unpaired t tests (*, p ≤ 0.05; **, p ≤ 0.01; ***, p ≤ 0.001; ns, not significant), except for E, in which a paired t test was used.