Extended Data Fig. 2: Corresponds to Fig. 2.

a. Graph shows % IL-9⁺ of d8 human Th9 cells restimulated with vehicle (n = 7), IL-1β (n = 3), IL-18 (n = 4), IL-33 (n = 4), IL-36α (n = 7), or IL-36γ (n = 7). b, c. Timelines show experimental design; graphs shows % IL-9⁺ (n = 6) for human non-restimulated T cells differentiated as in Fig. 1a, with STAT5 inhibitor or STAT6 inhibitor from d0-5 (b) or d5-8 (c). d–f. Bar graph shows concentration (pg/mL, ELISA) of IL-2 (d), IL-4 (e), and IL-9 (f) in supernatants of human in vitro differentiated Th1- or Th9 (d5), or Th2 (d14) cells, after 48 h restimulation. g, h. d8 human Th9 cells were restimulated with escalating doses of IL-2 (b) (2, 20, or 200 ng/mL) or IL-4 (c) (6, 60, or 500 ng/mL). *p < 0.05, paired t-test (n = 3). i, j. Representative flow plots (i) and bar graph (j) show % IL-9⁺ cells in resting (d4) murine Th9 cells restimulated with IL-2 + IL-4 (n = 10), **p < 0.01, Wilcoxon test. k. Bar graph shows pooled results d8 human Th9 cells restimulated with vehicle (n = 9), IL-6 (n = 3), IL-7 (n = 5), IL-12 (n = 3), IL-21 (n = 3), or TSLP (n = 4). l, m. Bar graphs show pooled (n = 6) mean fluorescence intensity (MFI) of pSTAT5 (l) and pSTAT6 (m) in d8 human IL-9⁺ and IL-9⁻ Th9 cells restimulated with IL-2, IL-4, or IL-2 + IL-4. *p < 0.05, **p < 0.01, Wilcoxon. n. Bar graphs show pooled (n = 3) % IL-9⁺ d8 human Th9 cells stimulated with IL-2 and IL-4 in the presence of STAT5 or STAT6 inhibitor. *p < 0.05, paired t-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 are 2-sided.

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