Fig. 2: SETD5 inhibits H3K27 acetylation and enhancer activation of adipogenic genes.

a Heatmap representation of H3K27 acetylation during 3T3-L1 adipogenesis. Heatmap shows ChIP-seq data for H3K27ac at a 20-kb region centered on H3K27ac enriched region in 3T3-L1 preadipocytes transduced with empty virus or SETD5 at the indicated time after MDI induction. There were 11,033 H3K27ac enriched regions that were induced by differentiation stimuli (48 h) in control preadipocytes. These regions were classified into distal enhancer (9,546 regions) and promoter (1,487 regions) based on the absence and presence of H3K4me3 signals. H3K27ac enriched regions were further classified into repressed (<0.5-fold), induced (>2-fold), and unchanged by SETD5 transduction. b, c Gene ontology (GO) analysis (b) and HOMER motif analysis (c) of differentiation-induced and SETD5-repressed H3K27ac enriched enhancer regions. p-values of motif analysis were calculated as one-tailed. d Venn diagram and heatmap representation of differentiation-induced and H3K27ac enriched regions and C/EBPβ and C/EBPδ binding regions. 443 regions were bound by C/EBPβ, 71 regions were bound by C/EBPδ, and 935 regions were bound by both. Heatmap shows ChIP-seq data for H3K27ac, C/EBPβ, and C/EBPδ at a 20-kb region centered on H3K27ac enriched region. e Genome browser representation for H3K27ac, C/EBPβ, and C/EBPδ on adipogenic genes. Pparg gene has two transcription start sites (TSS). Tracks 1, 2, and 7 are the same data as Supplementary Fig. 1d. f, g ChIP-qPCR analysis of H3K27ac on Cebpa gene during adipogenesis. 3T3-L1 preadipocytes transduced with empty virus or SETD5 were subjected to ChIP-qPCR analysis of H3K27ac at the indicated time after MDI induction of differentiation (f). 3T3-L1 preadipocytes transfected with siRNA targeted to Setd5 were subjected to ChIP-qPCR analysis of H3K27ac at the indicated time after Dex induction of differentiation (g). d, e ChIP-seq for C/EBPβ and C/EBPδ was from the previously published paper¹⁹. f Data are mean ± SEM (Empty n = 3; SETD5 n = 4; independent experiments). g Representative of two independent experiments. Data mean ± SD of three technical replicates. f, g One-way ANOVA with Tukey’s multiple comparisons test. **p < 0.01. Source data are provided as a Source data file.