Extended Data Fig. 5: Corresponds to Fig. 5.

a–c. Pooled counts of pulmonary CD45⁺TCRβ⁺CD4⁺CD44⁺IL-13⁺ cells (a, n = 8, PBS; n = 13, papain), CD45⁺TCRβ⁺CD4⁺CD44⁺IL-17A⁺ cells (b, n = 3, PBS; n = 6, papain), and CD45⁺TCRβ⁺CD4⁺CD44⁺IL-2⁺ cells (c, n = 8, PBS; n = 13, papain) from mice treated with isotype versus αMHC2 between papain sensitization and challenge. d. Th9 cells were differentiated in the presence of vehicle versus BMS-509744 (Itk inhibitor; 0.5 μM, 1 μM, 1.5 μM) and restimulated with αCD3 + αCD28 (n = 3) e-f. Bar graphs (n = 7) show %IL-9⁺ (e) of Th9 and %IL-4⁺ (f) of Th2 cells restimulated with αCD3 + αCD28 and vehicle versus BMS-509744 (0.5 μM, 1 μM), *P < 0.05 paired t-test. g. Design of inducible Itk-deleted mouse. Itk^flox/flox were crossed ERT-Cre mice to generate Itk^ERT mice. h. Representative flow plot and bar graph show %IL-9⁺ cells of Th9 cells from TAM-treated (Itk-deleted, n = 3) versus TAM-untreated (n = 5) mice, *P < 0.05 Mann–Whitney U-test. i. Timeline shows model of papain-induced airway inflammation with Itk deletion. Mice were treated with vehicle versus tamoxifen between sensitization and challenge. j. Gel shows that injection of tamoxifen results in deletion of WT Itk gene in mice from (i). k–n. Representative periodic acid-Schiff (PAS) stained images (k), pooled histology scores (l, n = 3, PBS; n = 5, papain), pulmonary CD45⁺TCRβ⁺CD4⁺CD44⁺IL-9⁺ (Th9) (m, n = 5, PBS; n = 10, papain), and pulmonary CD45⁺TCRβ⁺CD4⁺CD44⁺IL-13⁺ (Th2) cell counts (n, n = 3, PBS; n = 5, papain), from Itk^f/f treated with vehicle versus tamoxifen as in (i). o–r. Pooled counts of pulmonary CD45⁺CD4^- IL-9⁺ cells (o, n = 5, PBS; n = 8, papain), CD45⁺TCRβ⁺CD4⁺CD44⁺IL-13⁺ cells (Th2) (p, n = 5, PBS; n = 8, papain), CD45⁺TCRβ⁺CD4⁺CD44⁺IL-17A⁺ cells (Th17) (q, n = 4, PBS; n = 7, papain), and CD45⁺TCRβ⁺CD4⁺CD44⁺IL-2⁺ cells (r n = 4, PBS; n = 7, papain) ILC2-deficient mice treated with isotype versus αMHC2 between papain sensitization and challenge. For all in vivo experiments, *P < 0.05, **P < 0.01, ***P < 0.005, ****P < 0.001, Mann–Whitney U-test. For all line/bar graphs, error bars show ± s.e.m. Box plots show all data points (min to max, lines at median). All statistical tests were two-sided.

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