Fig. 5: Eaf3 interacts with Sin3 to maintain histone hypoacetylation in the Ash1-H3K36me2-marked and Eaf3-occupied regions.

a Y2H assays between Eaf3 and Sin3. b Co-IP assays were performed with the strains co-expressing SIN3-GFP and EAF3-Flag. Two independent experiments had similar results. c Relative abundance of H3K4ac, H3K27ac, and H4K5ac in the indicated strains. Ratios of H3K4ac, H3K27ac, and H4K5ac to H3 in western blots were calculated by ImageJ software respectively and that of WT was set as 1. Values are means ± SD from five biological replicates. d, e Metagene plots and heatmaps showing H3K27ac ChIP-seq signals of the Eaf3-occupied regions in the WT and ∆eaf3 strains. f A Venn diagram showing significant overlap of gene sets between H3K27 hyperacetylated and Eaf3-occupied. P-values were calculated by two-sided Fisher’s exact test. g Box plots and violin plots showing average transcription of the genes with and not Eaf3 occupancy in the indicated strains. h Box plots and violin plots showing average transcription levels of the genes with and not Ash1-H3K36me2 in the indicated strains. For box-plot, the horizontal lines from top to bottom represent the maximum, first quartile, median, third quartile, and minimum of the total data, respectively. i IGV showing representative genes with Eaf3-GFP, H3K36me2, and H3K27ac occupancy from ChIP-seq assay and RNA-seq in the indicated strains. ChIP-qPCR assays were performed to validate the ChIP-seq results. Relative enrichments of Eaf3 (l), H3K36me2 (m), and H3K27ac (n) are shown. I and II indicated the amplified fragments in the ChIP-qPCR assays. Values are means ± SD from three technical replicates. For c, g, h, j–l, statistical significance was assessed using a two-tailed Student’s test (**P < 0.01). The exact P-values and western blots are shown in the Source Data. ns, no significance. For d, e, i, metaplots, heatmaps and IGV snapshots are shown with one biological replicate, and similar results were gained in other replicates.