Figure 1

A subset of intraepithelial lymphocyte precursors (IELPs) expresses T-bet. (a,b) Flow cytometry analysis of cells purified from the epithelium of the (a) small intestine or the (b) thymus of 4-week-old C57BL/6 mice or TBGR mice. Pregating is shown in Supplementary Figure S1a,b. Histogram data of IELs are representative of at least three independent experiments. Numbers denote percentage of cells in the gate (mean±s.e.m., at least 3 mice per genotype for thymi). (c) Tbx21 mRNA expression of different flow cytometry-sorted thymocyte populations from TBGR mice analyzed by qPCR, gating strategy for ZsGreen subpopulations as shown in b (double positive (DP); single positive (SP)). (d,e) Immunofluorescent staining of sections from the (d) small intestine or the (e) thymus of TBGR mice for the indicated markers. Representative pictures taken with (d) × 20 or (e) × 10 original magnification are shown. (f) Flow cytometry analysis of H2-K^b, CD122, Qa2, and CD24 in relation to ZsGreen (T-bet) in IELPs from TBGR mice. (g) Flow cytometry analysis of thymic maturation markers CD24 and H2-K^b in ZsGreen⁻, ZsGreen^int, and ZsGreen^high IELPs. Numbers in quadrant gates denote percentages (mean±s.e.m.) from four analyzed TBGR mice. (h) Contour plot of IELPs analyzed for the expression of T-bet vs. Helios and Nur77, respectively. Numbers in quadrant gates denote percentages (mean±s.e.m.) from four analyzed TBGR mice. (i,j) The 6–8-week-old TBGR mice were treated with 5 μg ml⁻¹ FTY720 for 2 weeks or left untreated as a control. After treatment IELPs were analyzed for the expression of (i) T-bet vs. H2-K^b, (j) T-bet vs. Qa2, and (j) CD24 vs. Qa2. Numbers in quadrant gates denote percentages (mean±s.e.m.) from at least three analyzed TBGR mice per treatment group. (k) Flow cytometry analysis of ZsGreen, CD24, and H2-K^b in IELPs from wild-type TBGR, Tbx21^−/− TBGR, and Il15^−/− TBGR mice. Numbers in quadrant gates denote percentages (mean±s.e.m.) from at least three mice per genotype.

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