Figure 2: Transcriptional co-regulation is dependent on Set1 and Jhd2 catalytic activities and H3K4 methylation. | Nature Communications

Figure 2: Transcriptional co-regulation is dependent on Set1 and Jhd2 catalytic activities and H3K4 methylation.

From: Counteracting H3K4 methylation modulators Set1 and Jhd2 co-regulate chromatin dynamics and gene transcription

Figure 2

(a) Western blots for H3K4me1, H3K4me2 and H3K4me3 in WT, deletion mutants (set1Δ, jhd2Δ and set1Δ jhd2Δ), catalytic-dead mutants (set1-C1019A and jhd2-H427A) and Jhd2 overexpressing strain (ADH1pro-JHD2) are shown. Histone H3 serves as loading control. Twofold serial dilutions of cell extracts from the indicated strains were applied in lanes 5–16. Graph shows quantitation of western blots using densitometry. Fold-change in a H3K4 methyl mark level normalized to the total H3 level in a mutant is shown relative to that in control WT (set as 1). (b) Fold-change in PHO89 and LCP5 sense transcript levels in the indicated mutants relative to WT are shown. For (b,c), error bars denote ±s.e.m. from four independent experiments (n=4). Statistical significance calculated using the Student’s t-test. PHO89: *P-value <0.05, **P-value <0.005, ***P-value <10−5; LCP5: *P-value ≤0.005. (c) Fold-change in SER3 and CHA1 transcript levels in the indicated mutants relative to WT are shown. SER3: *P-value <10−5, CHA1: *P-value <10−3. (d) Western blots for bulk H3K4 methylation and H3 levels in WT strain or strains overexpressing SET1 or hyperactive SET1-G990E. Graph shows quantitation of western blots using densitometry. (e) Four-way Venn diagrams showing the number of shared up- or down-regulated sense transcripts in each combination of indicated deletion or overexpression mutant strains. The total number of up- or down-regulated transcripts in each mutant is indicated in parentheses. For (a–d), see Supplementary Figs 20 and 21 for full-length image of blots.

Back to article page