Fig. 4. TGF-βRII is required for mature T_H17 cells to produce IL-10.

a Flow cytometric analysis of in vitro cultured total CD4⁺ T cells. Cells were polarized under T_H17 condition for 5 days. A pre-gate on viable (DAPI⁻), CD4⁺ T cell population is applied. b Statistical analysis of IL-10⁺ T_H17 and T_R1^exTH17 cells is shown by one representative experiment out of three. Each dot represents one mouse (n_{wild type} = 3, n_KO = 5). Mean ± S.D.; *P < 0.05, ***P < 0.001 by Welch’s t-test. c Flow cytometric analysis of small intestinal CD4⁺ T cells after anti-CD3 mAb treatment. The indicated reporter mouse lines were used. Top panels are pre-gated on Foxp3⁻ CD4⁺ T cells. Bottom panels are gated on YFP⁺ cells, as shown, and the indicated populations are identified on the basis of the reporter molecules as follows: T_R1^exTH17 cells: IL-10^eGFP+ IL-17a^Kata−; IL-10⁺ T_H17 cells: IL-10^eGFP+ IL-17a^Kata+; T_H17 cells: IL-10^eGFP– IL-17a^Kata+. d, e Frequencies (d) and numbers (e) of the indicated populations are shown. One representative experiment out of three is shown. Each dot represents one mouse (n_{wild type} = 5, n_KO = 5). Mean ± S.D.; ns, not significant; *P < 0.05, **P < 0.01 by Mann–Whitney U test. f. Experimental design for Co-transfer of total CD4⁺ T cells isolated from wild-type Fate⁺ mouse and Tgfbr2^Flox/Flox Fate⁺ mouse lines. g. Flow cytometric analysis of small intestinal co-transferred CD4⁺ T cells after anti-CD3 mAb treatment. Small intestinal lymphocytes were isolated from the indicated mouse lines and intracellular staining was then performed to identify the indicated cell populations in combination with reporter molecules for IL-10 (eGFP) and IL-17a lineage (YFP). All cell populations are pre-gated on CD4⁺ T cells. h Frequencies of T_R1^exTH17 and IL-10⁺ T_H17 in G is shown by one representative experiment out of two. Each dot represents one recipient mouse (n = 3). Mean ± S.D.; *P < 0.05, ***P < 0.001 by Welch’s t-test. Source data are provided as a Source data file.