Extended Data Fig. 9: RUNX–CBFβ regulates the development of cDCs.
From: A RUNX–CBFβ-driven enhancer directs the Irf8 dose-dependent lineage choice between DCs and monocytes
a, Genome Browse images of RUNX1 and RUNX2 ChIP-seq tags in HSPC cell lines around the Irf8 gene and enhancers, retrieved from previous reports39,40,41. ATAC-seq data on WT MDPs newly obtained in this study (n = 2 biologically independent samples, each using 1 mouse) and H3K27ac ChIP-seq data on WT MDPs retrieved from our previous publication21 (n = 2 biologically independent samples, each using 20 mice) are shown for reference. b,c, WT LSK cells were transduced with a lentivirus encoding shRNA against Cbfb and cultured with SCF and Flt3L as in Fig. 7. Representative FACS plots on day 7 are shown in (b). The percentages of MDPs and CDPs (day 5, n = 3 cell cultures), cDCs, pDCs, and monocytes/macrophages (day 7, n = 4 cell cultures) among GFP+ cells are shown in (c). Data are representative of two independent experiments, which yielded similar results. The bar graphs are shown as mean + SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed Student’s t test). The exact P values are provided in Source Data. d, Representative histograms of reporter assays in MDPs shown in Fig. 7f. e, mRNA expression of Runx1, Runx2, Runx3, and Cbfb analyzed by RNA-seq in the indicated cell types from WT mice (n = 2 biologically independent samples). Data are shown as mean. f, Normalized microarray intensities of Runx1, Runx2, Runx3, Cbfb, and Irf8 in IRF8– and IRF8+ LMPPs (n = 2 biologically independent samples). Data are shown as mean. g, A Genome Browser image of sequence conservation at the human IRF8 gene locus against the mouse genome. The region corresponding to mouse +56 kb Irf8 enhancer (dotted line) and the region used for the +56 kb enhancer reporter assay (orange) are indicated. Positions of the two RUNX motifs are shown as blue arrow lines. Mac, macrophage; NC, negative control.
