Fig. 3

ILC2s give rise to IL-17 producing cells at a clonal level. a–d Single ILC2s from CRS NP were sorted by FACS into 96-well round bottom plates pre-seeded with OP9-DL1 and stimulated with IL-2 (20 U/mL), IL-7, IL-1β, IL-23, and TGF-β (20 ng/mL each). a After 14-21 days, cultures were analyzed for intracellular cytokine production (IL-5, IL-13, and IL-17A) after PMA/ionomycin restimulation. b Representative flow cytometric analysis of intracellular cytokine production of three clones. c Quantification of concentration of cytokines of three clones. d RORC expression in IL-17A⁺ and IL-17A⁻ clones (n = 8 per group). Each symbol represents an individual clone and the horizontal lines represent the mean. e, f Clonal cultures were resorted and cultured for 5 days on OP9-DL1 with IL-2, IL-7, IL-1β, IL-23, and TGF-β; or IL-2, IL-7, IL-33, and TSLP. e Quantification of intracellular IL-5, IL-13, and IL-17A production after PMA/ionomycin stimulation (n = 7–8 clones per group). Each symbol represents an individual donor and the horizontal lines represent the mean. f Representative flow cytometric analysis of cytokine production of an IL-17⁺ clone. Data are presented as individual values (c–e) of three independent cloning experiments with one donor each. *p < 0.05, **p < 0.01, ***p < 0.001 as determined by one way ANOVA or Student’s t test. Source data