Histone modification in dendritic cell progenitors and dendritic cells

To understand the roles of Irf8 3' enhancers on Irf8 expression during cDC1 differentiation, we performed Cleavage Under Targets and Tagmentation (CUT&Tag) analysis for H3K27ac of bone marrow cells and splenocytes from wild-type, and the Irf8 enhancer-deficient mice. Phenotypic analysis and integrated analysis of CUT&Tag data with transcriptome data of these mouse mononuclear phagocytic lineage cells revealed that the +56 kb enhancer acts on other 3' enhancers via an IRF8-dependent TF program and the +32 kb enhancer acts in cis to activate other 3' enhancers. Overall design: Murine type1 classical dendritic cells (cDC1s), cDC1 subset-committed progenitors (pre-cDC1s), common DC progenitors (CDPs), and monocyte-DC progenitors (MDPs) were isolated from WT, +56 kb enhancer-heterozygous deficient (d+56het), or +32 kb enhancer-heterozygous deficient (d+32het). The IRF8-rescued cDC1s were prepared from Lin– cells from WT, +56 kb enhancer-deficient (d+56), or +32 kb enhancer-deficient (d+32). These cells were subjected to CUT&Tag analysis (biological triplicates for each cell type).