Extended Data Fig. 2: Runx1 and Runx3 heterodimerization and cell intrinsic increase of ILC2 progenitor.
(a) Venn diagram showing the H3K4me3 regions in the whole genome between Rx3+/+ and Rx3+/W mice. (b) Graphs showing numbers of lymph nodes (LNs) at the indicated location and Peyer’s patches (PPs) in Rx3+/+(n = 8 for LNs and n = 6 for PPs), Rx3+/W(n = 8 for LNs and n = 6 for PPs), and Rx3W/W (n = 8 for LNs and n = 6 for PPs) mice. Mean ± SD. ****: p < 0.0001 (ANOVA with Tukey’s multiple comparisons test for LNs and unpaired student t-test,two-sided for PPs). (c, d) Schemas showing the structure of the Rx1TY1 allele with amino acid sequences of theTY1 tag (c) and the Rx3Flag allele with amino acid sequences of the Flag tag (d) generated by CRISPR/Cas9 genome editing. Images of agarose gel electrophoresis of one-week-old F0 founders are shown at the bottom. A single experiment was performed. (e) Scheme showing the principle of AlphaScreen to examine the interaction of biotin-tagged Runx with Flag-tagged Runx. (f) Effect of Cbfβ and different oligonucleotides with or without the canonical Runx recognition motif (5′-TGTGGT-3′and 5′-ACCACA-3′) on the interaction between biotin-tagged Runx1 and Flag-tagged Runx3 in AlphaScreen. (g) Scheme showing the experimental flow of the bone marrow chimera experiment. The right graph shows a summary of the frequencies of CD45.1+ and CD45.1+CD45.2+ ILC1, ILC2P, ILC2, and ILC3 in the bone marrow of three recipient mice (n = 3). Mean ± SEM. *: p < 0.05, ****: p < 0.0001 (unpaired student t-test, two-sided for ILC2 and Welch’s t-test, two-sided for ILC2P).
