Fig. 2: Pulmonary ILCs shift from ILC2 to ILC1s as silicosis progresses.

a Experimental schematic. Saline or CS was administered to C57BL6J mice that were sacrificed at time 1: 8–10 weeks or time 2: 12–14 weeks post treatments. b Representative Masson’s trichrome staining in lung section images showing whole lung lobes and zoomed in areas. Scale bar = 600 μm (upper panel), 200 μm (middle panels), and 50 μm (lower panels). The bar graph compares fibrotic area percentages in lung sections (n = 5 biological replicates). c Western blot showing fibronectin (FN) and collagen I (Col1) expression in CS-treated mouse lung groups. β-Actin was used as a loading control. Uncropped blots are in source data. The bar graph compares relative protein expression in lungs from different group (n = 4 biological replicates). d Flow plots showing NKp46⁺ (ILC1) and ST-2⁺ (ILC2) percentages in ILCs at indicated times. Bar graphs compare NKp46⁺ and ST-2⁺ percentages in ILCs and NKp46⁺/ST-2⁺ ILC ratios in lungs (n = 8 biological replicates). e Flow plots showing T-bet⁺ and Gata-3⁺ percentages in ILCs at indicated times. Bar graphs compare T-bet⁺ and Gata-3⁺ percentages in ILCs and T-bet⁺/Gata-3⁺ ILC ratios in lungs (n = 8 biological replicates). f Experimental schematic design: Saline or CS was administered to Rag1^–/– mice that were sacrificed at indicated time points. g Flow plots showing NKp46⁺ (ILC1) and ST-2⁺ (ILC2) percentages in ILCs at indicated times. Bar graphs compare NKp46⁺ and ST-2⁺ percentages in ILCs and NKp46⁺/ST-2⁺ ILC ratios in lungs (n = 5 biological replicates in the saline group, n = 7 biological replicates CS 8-10 week, n = 8 biological replicates CS 12–14 weeks). h Flow plots showing T-bet⁺ and Gata-3⁺ percentages in ILCs at indicated time. Bar graphs compare T-bet⁺ and Gata-3⁺ percentages in ILCs and the T-bet⁺/Gata-3⁺ ratio in lungs (n = 6 biological replicates). i Western blot comparing fibronectin (FN) and collagen I (Col1) expressions in CS-treated C57BL/6 J and Rag1^–/– mice lungs 12–14 weeks after CS treatment. β-Actin was used as a loading control. Uncropped blots are in source data. The bar graph compares relative protein expressions from different group (n = 4 biological replicates). j Bar graphs compare ratios of T-bet⁺/Gata3⁺ ILC in C57BL/6J and Rag1^–/– mice lungs 12–14 weeks after CS treatment (n = 8 biological replicates in C57BL/6 group, n = 6 biological replicates in Rag1^–/– group). k Schematic depicting bleomycin (BLM)-induced pulmonary fibrosis: BLM was administered to C57BL6J mice twice, at weeks 0 and 1. Mice were sacrificed at time 1: week 3 or time 2: week 5 post the last BLM treatment. l Flow plots showing NKp46⁺ (ILC1) and ST-2⁺ (ILC2) percentages in ILCs at indicated time points. Bar graphs compare NKp46⁺ and ST-2⁺ percentages in total ILCs and the NKp46⁺/ST-2⁺ ILC ratio in lungs (n = 8 biological replicates). m Flow plot showing ST-2 and IL-18Rα expression levels in ILCs from B6 mouse silicotic lungs. Bar graphs compare IL-18Rα⁺ST-2⁺ ILC percentages with silicosis progression (n = 8 biological replicates). Correlated ST2 and IL-18Rα expression (n) and correlated NKp46 and ST-2 expression levels (o) on ILCs from CS-treated B6 mice (n = 12 biological replicates). p Flow plots showing cell retention marker expression on IL-18Rα⁺ST-2⁺ ILCs in silicotic lungs at 12–14 weeks after CS treatment. Flow plots and bar graphs were representative of four biologically independent experiments with similar results. Individual mice are plotted on graphs. Data are shown as the mean ± SEM. P values were calculated using two-tailed unpaired Student’s t-tests (two groups) or one-way ANOVA followed by Tukey’s tests (three groups) and indicated on graphs. Source data are provided in a source data file.