Fig. 2

Id3 regulates the pool size of NK1.1⁻ IELPs. a, b Flow cytometric analysis of thymocytes of 6–12-week-old Id3^Gfp/+ (littermate controls) and Id3^Gfp/Gfp mice. Cells were pre-gated on NK1.1⁻ IELPs as shown in Supplementary Fig. 1a and analyzed for Id3 (GFP) expression vs. CD122 (a) and T-bet (b). Bar diagram represents percentage (Mean ± SEM) of NK1.1⁻ IELP subpopulations based on CD122 and T-bet expression as in a, b. d Contour plot shows GFP vs. CD24 expression in thymic NK1.1⁻ IELP. e Bar diagram shows GFP MFI from d. f Contour plot shows GFP vs. Nur77 expression in thymic NK1.1⁻ IELPs. g Bar diagram shows percentage (Mean ± SEM) of the indicated populations as gated in f. h Upper row shows flow cytometric analysis of lymphocytes isolated from the epithelium of the small intestine of 6–12-week-old Id3^Gfp/Gfp mice with Id3^Gfp/+ littermates as controls. Cells were pre-gated on DAPI⁻ CD45⁺ CD4⁻. Lower row shows further flow cytometric analysis of TCRβ⁺ IELs for CD8β⁺ and CD8α⁺. Numbers denote the percentage of cells in the gate (Mean ± SEM). i Bar diagrams represent percentages (upper, Mean ± SEM) and absolute cell numbers (lower, Mean ± SEM) of TCRβ⁺ and TCRγδ⁺ IELs as shown in h. j Dot plot shows GFP expression of small intestine TCRβ⁺ IELs gated as in h. a–j Experiments were performed twice with at least six mice per genotype. k Bar diagram represents mRNA expression of Id2 for different thymocyte populations from three 6–12-week-old TBGR mice performed by qPCR. l Flow cytometric analysis of thymocytes of 4–8-week-old Id2^Gfp/+ mice. Dot plot shows the expression of CD24 vs. H2-K^b maturation of thymic NK1.1⁻ IELPs and numbers denote the percentage of cells in the gate (Mean ± SEM) from four mice. Histogram represents MFI expression of GFP (Id2) of the indicated thymic NK1.1⁻ IELP populations for Id2^Gfp/+ and Id2^+/+ mice as controls. m Histogram represent MFI expression of GFP (Id2) of the indicated IEL populations in the small intestine for Id2^Gfp/+ and Id2^+/+ mice from one representative mouse. n Bar diagram shows MFI (Mean ± SEM) of GFP (Id2) in thymic NK1.1⁻ IELPs and small intestine IELs as shown in l, m from four mice