Extended Data Figure 6. Zbtb46⁺ ILC3s and cDCs are distinct cell lineages.

a. Representative flow cytometry plots showing the YFP⁺ cells in cDCs and CCR6⁺ ILC3s from the large intestine of indicated fate-mapping mice. b. Schematic of progenitor cell transfer. pre-DC or CHILP were sort-purified from the bone marrow of Zbtb46^GFP/+ and transferred into Rag2^−/− Il2rg^−/− recipient mice. Six weeks later, ILC3s and Zbtb46-GFP expression were analyzed in the intestine of recipient mice. PBS injection was used as a negative control. pre-DCs were gated as: (Ter119, CD19, B220, CD3, CD5, NK1.1, Ly6G, CD11b)⁻CD127⁻MHCII⁻CD117⁻CD16/32⁻CD11c⁺Flt3⁺GFP⁺; CHILP were gated as (Ter119, B220, CD19, NK1.1, CD3, CD5, CD11b, CD11c, Ly6G)⁻CD45⁺CD127⁺α4β7⁺Flt3⁻CD25⁻CD117⁺. c. Representative flow plots of recipient mice injected with indicated progenitor cells. γc staining was used to differentiate the host-derived and recipient-derived ILCs. d. Gating strategies for progenitors and LTi cells from the fetal tissue. e. RORγt (RORE) motif analysis of the Zbtb46 locus in ATAC-seq data from CCR6⁺ ILC3s. f. Zbtb46 expression relative to Hprt in sort purified CCR6⁺ ILC3s with or without exposure to the RORγt inhibitor GSK805 (n = 11 mice). g. Quantification of Zbtb46 MFI in CCR6⁺ or CCR6⁻ ILC3s from the large intestine of indicated mice (n = 6 or 5 mice per group for both subsets). h. Quantification of cell numbers of Zbtb46-GFP⁺ cells in indicated ILC3 subsets from the large intestine of water or ABX-treated Zbtb46^GFP/+ mice (n = 5 mice for all subsets). i. Correlation analysis of ZBTB46 expression and disease scores in Ulcerative colitis patients. Data are pooled from three or two individual experiments in f and g, data are representative of two independent experiments in h. Data are shown as means ± S.E.M in g and h. Statistics are calculated by two-tailed paired Student’s t-test in f, two-tailed unpaired Student’s t-test in g and h, and the correlation is determined by simple linear regression in i.