EILPs differentiate into ILCps. ALPs, EILPs, and ILCps were isolated from Tcf7^EGFP/+ mice by cell sorting and cultured for 4 d. (A–D) Cultures were supplemented with IL-7. Data are representative of three independent experiments. ***, P < 0.005. (A) ICOS and NK1.1 expression analyzed by flow cytometry. (B) Quantification of total cells. (C) GFP, Mac-1, and Thy1 expression of ICOS⁻ NK1.1⁻ cells from A. (A and C) Numbers indicate the percentage of cells in each gate. (D) Quantification of Mac-1⁺ cells from C. (B and D) Data are presented as mean ± SD for triplicate wells. A two-tailed unpaired Student’s t test was performed to determine significance. (E) Flow cytometric analysis of IL-7Rα expression on ex vivo EILPs or NK1.1⁻ ICOS⁻ Tcf7⁺ Thy1⁺ cells from EILPs and ILCps cultured without IL-7. (F) Flow cytometric analysis of PLZF expression measured by intracellular staining by ex vivo EILPs, in vitro EILP-derived NK1.1⁻ TCF-1⁺ cells, and ILCp-derived NK1.1⁻ TCF-1⁺ cells cultured with IL-7. (E and F) Data are representative of two independent experiments.