Fig. 5
TRIM33 modulated the PU.1 transcriptional function and Irf8 expression via chromatin occupancy. A The interaction of TRIM33 with CDK9 in Mutu DC1940 cells was detected using coimmunoprecipitation. B, C Venn diagrams indicating the intersection between genomic regions bound by the indicated factors in CDPs. CDP TRIM33, CDK9, and S2 Pol II peaks were called from CUT&Tag data. CDP ATAC-seq and H3K27ac-ChIP region data were retrieved from the GSE132240 and GSE132239 datasets. D ChIP-Atlas analysis of RNA-seq-detected progenitor DEGs following TRIM33 deletion. FE, fold enrichment. E Intersection between TRIM33-reliant and PU.1-reliant transcriptomes. The percentages of TRIM33-dependent/repressed genes under the control of PU.1 are shown. F Intersection between CDP TRIM33 CUT&Tag peaks and cDC1 IRF8-ChIP (GSE66899) peaks. G Analysis of shared TRIM33 and PU.1 peaks in CDPs. PU.1-ChIP peaks were called from GSE57563. The superenhancer analysis was performed with SEA v3.0. H–L Visualization of TRIM33, CDK9, S2 Pol II CUT&Tag, PU.1-ChIP, H3K27ac ChIP, and ATAC-seq data at the indicated loci in CDPs. The reported +32 kb and +41 kb enhancer sites of Irf8 are shaded in orange. ChIP‒qPCR analysis of TRIM33 (M), CDK9 (N), S2 Pol II (O), and total Pol II (P) occupancy at different regions of the Irf8 gene in BM Lin– cells from tamoxifen-treated Trim33fl/fl (WT) and Trim33fl/fl Cre-ERT2 (KO) mice. N = 3–4. The data are representative of 2 independent experiments. Bars represent the means ± SEMs. The data were analyzed using two-way ANOVA followed by Bonferroni’s multiple comparison test. *P < 0.05, **P < 0.01, ***P < 0.001, and ****P < 0.0001
