Differentiation capacity of KLRG1⁺ ILCs and NKp46⁺ ILCs. (A) Quantification of relative GATA3 and RORC expression as compared with GAPDH in different ILC subsets isolated from tonsils and PB. (B) Representative histograms of GATA3 and RORγt expression in KLRG1⁺ ILCs and NKp46⁺ ILCs, respectively, after culture for 5 d on OP9 or OP9-DL1 in the presence of IL-2 (20 U/ml) and IL-7 (20 ng/ml). (C) Quantification of GATA3 expression on KLRG1⁺ and NKp46⁺ ILCs after culture as in B. (D) Quantification of RORγt expression on KLRG1⁺ and NKp46⁺ ILCs after culture as in B. (E) Representative flow cytometric analysis of CRTH2, GATA3, NKp44, and RORγt expression on KLRG1⁺ ILCs, ILC2s, and NKp46⁺ ILCs after culture for 5 d on OP9-DL1 in the presence of IL-2 (20 U/ml) and IL-7 (20 ng/ml). (F) Quantification of the percentage of cells expressing CRTH2, NKp44, GATA3, RORγt, EOMES, and T-bet upon culture as in E. Each dot represents one individual donor. *, P < 0.05; **, P < 0.001; ***, P < 0.0001; ****, P < 0.00001 (one-way ANOVA). Data in B and E are representative of at least three donors from at least three independent experiments. AU, arbitrary units; CTRL, control; MFI, mean fluorescence intensity.