Fig. 5: SETD5 is co-recruited to primed enhancers with NCoR-HDAC3 complex and diminishes prior to enhancer activation.

a Venn diagram (left) and heatmap (right) representation of genomic regions occupied by SETD5-V5, NCoR, and HDAC3. Approximately 60% of NCoR and 71% of HDAC3 genomic binding regions were co-occupied by SETD5-V5. Heatmap analysis showing ChIP-seq data for SETD5-V5, NCoR, and HDAC3 at a 20-kb region centered on SETD5-V5 binding region. b Genome browser representation for SETD5-V5, NCoR, HDAC3, and H3K27ac on Cebpa and Pparg genes in 3T3-L1 preadipocytes. Tracks 1-2 and 7-9 are the same data as Fig. 4c. c, d ChIP-qPCR analyses of NCoR2 (c) and HDAC3 (d) on Cebpa and Pparg genes during the early adipogenesis. 3T3-L1 preadipocytes transduced with control empty virus were subjected to ChIP-qPCR analyses using anti-NCoR2 or anti-HDAC3 antibody. Data are mean ± SEM (0, 48 h n = 5; 6 h of NCoR2 n = 4; 6 h of HDAC3 n = 5; 12, 24 h n = 3; independent experiments). e ChIP-qPCR analysis of SETD5-V5 on Cebpa and Pparg genes under knockdown of ANAPC11 or CDC20. 3T3-L1 preadipocytes transduced with SETD5-V5 were transfected with control siRNA or siRNA targeting to Anapc11 or Cdc20 and subjected to ChIP-qPCR analysis at 6 h and 24 h of differentiation using anti-V5 antibody. ChIP signal was presented as relative SETD5 recruitment. Data are mean ± SD of four technical replicates. f Schematic model of SETD5-NCoR-HDAC3 complex formation and the loss of SETD5 on enhancers during the transition from primed to active states. Heatmap shows the degree of recruitment of SETD5, NCoR2, and HDAC3 to Cebpa and Pparg enhancers (based on Figs. 4d, 5c, and 5d). Darker color indicates more recruitment. a, b ChIP-seq for NCoR and HDAC3 was from the previously published paper³². c–e One-way ANOVA with Tukey’s multiple comparisons test. *p < 0.05; **p < 0.01. Source data are provided as a Source data file.