Extended Data Fig. 4: The Irf8 +56 kb region regulates Irf8 expression and cell fate.
From: A RUNX–CBFβ-driven enhancer directs the Irf8 dose-dependent lineage choice between DCs and monocytes
a, Representative FACS plots of pre-cDC1s, pre-cDC2s, pre-DCs, and Ly6C– monocytes analyzed in Fig. 2a. b, IFN-α and IFN-β production by pDCs isolated from WT or ∆+56 mice followed by overnight stimulation with poly(U) (1.0 µg/mL) or CpG-A (10 µM) (n = 3 mice per genotype). Data are representative of two independent experiments, which yielded similar results. c, In vitro culture of MDPs. Ten thousand MDPs from WT, ∆+56, and Irf8–/– were cultured with Flt3L for 5 days. Representative FACS plots of DC subsets (upper panels) and their absolute cell numbers (lower panels) produced in the culture are shown (total n = 4 cell cultures). The data were pooled from two independent experiments. d,e, Bone marrow chimera experiments. WT or ∆+56 HSPCs (c-Kit+, 3.0 × 105 cells) were transplanted into irradiated mice (CD45.1+) together with 2.0 × 105 competitor WT whole bone marrow cells (CD45.1+). Cells were analyzed 2 months after transplantation by FACS and RT-qPCR. In (d), absolute numbers of progenitor populations in bone marrow and differentiated cells in spleens derived from WT or ∆+56 donor cells are shown. The data were pooled from two independent experiments (total n = 6 mice per genotype). In (e), Irf8 mRNA expression in donor-derived bone marrow progenitor populations (total n = 3 mice per genotype). The data were pooled from two independent experiments. Data in b, c (lower panels), d and e are shown as mean + SD. * P < 0.05, ** P < 0.01, *** P < 0.001 (two-tailed Student’s t test) with a fold-change greater than 1.5 or less than 0.66. The exact P values are provided in Source Data. N.D., not detected in (b) and not determined in (e).
