Fig 2: IL-18Ra-expressing ILC differentiate into ILC1-like cells during Mtb infection(A) Representative dot plots showing the gating strategy used for ILC subsets in the lungs of Mtb-infected C57BL/6 mice (top graphs): ILC1 (dark blue), ILC3 (red), IL-18Ra- ILC2 (green), IL-18Ra+ ILC (yellow), and T-bet+IL-18Ra+ ILC (light blue).(B) Unsupervised t-distributed stochastic neighbor embedding (t-SNE) distribution of total lung Lin-CD90.2+ populations at steady state (left graph) and during Mtb infection (right graph). Based on the gating strategy defined in (A), ILC subsets were depicted with the same color code.(C) Expression of GATA3 (MFI), ST2 (%), and Arg1 (%) in indicated ILC subsets at day 28 post-infection in C57BL/6 mice.(D) As in (C), but for the expression of T-bet (MFI) and CD49a (%).(E) Percentages of NKp46+ (left) and Eomes+ (right) cells in ILC1-like cells compared to in NK cells and ILC1 at day 28 post-infection.(F) Percentages of IFN-?, IL-5, and IL-17A-positive cells in the indicated ILC subsets after phorbol myristate acetate (PMA)/ionomycin stimulation at day 28 post-infection in C57BL/6 mice.(G and H) Absolute numbers of ILC1-like cells at the indicated days after Mtb infection (G) and absolute numbers of IL-18Ra+ ILC at the indicated days after Mtb infection (H). Before sacrifice, mice were injected with fluorescent anti-CD45.2 to distinguish vascular and parenchymal cells. ILC1-like cells and IL-18Ra+ ILC have been gated on lung-resident cells.(I) Percentages of IFN-?+ cells in the indicated ILC subsets after ex vivo stimulation with IL-12+IL-18 or not at day 28 post-infection in C57BL/6 mice.(J) Experimental settings for the adoptive transfer of ST2+IL-18Ra- and ST2+IL-18Ra+ ILC into Rag2-/-?c-/- before Mtb infection.(K) Percentage of ILC (Lin-CD45.2+CD90.2+CD127+) in the lung at day 21 post-infection in Rag2-/-?c-/- after adoptive transfer of ST2+IL-18Ra- ILC (green) and ST2+IL-18Ra+ ILC (yellow).(L) As in (J), but for T-bet expression in ILCs.In (C), (D), (G) and (H), data are representative of 5 independent experiments; in (E), (F), and (I), data are representative of 2 independent experiments; and in (K) and (L), data are a pool of 2 independent experiments, with each symbol representing an individual mouse. Graphs depict data as means ±SEMs, and statistical analysis was performed using 2-way ANOVA (F and I), 1-way ANOVA (C–E and G–H) or the Mann-Whitney test (K and L). *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001).