Fig. 5: Setd2 supports the reciprocal relationship of GATA3 and ST2 expression in tTreg cells.

a–h Splenocytes (SP) (a–d) and large intestinal (LI) LPLs (e–h) were isolated from 1-week (1w), 2-week (2w), or 3-week (3w)-old Foxp3^Cre-YFPSetd2^f/+ or Foxp3^Cre-YFPSetd2^f/f mice. (1w: n = 3 per group; 2w: n = 6 per group; 3w: Ctrl n = 7; KO n = 6). a, e Representative FACS plot of ST2 and GATA3 expression gated on Helios⁺ Treg cells. Percentages of GATA3⁻ST2⁺ cells (b, f), GATA3⁺ST2⁺ cells (c, g), and GATA3⁺ST2⁻ cells (d, h) in Helios⁺ Treg cells (CD3⁺CD4⁺Foxp3⁺Helios⁺) were analyzed by flow cytometry and shown. i–m Splenic (SP) Treg cells (CD3⁺CD4⁺YFP⁺) were sorted from mice of indicated genotypes and cultured with α−CD3/α−CD28 (TCR) with or without supplement of IL-2 (TCR + IL-2) or IL-33 (TCR + IL-33) for 24 h. i Expression of ST2 and GATA3 gated on Treg cells (CD3⁺CD4⁺Foxp3⁺) was analyzed by flow cytometry. j, k Percentage of GATA3⁺ cells (j) (TCR: n = 8 per group; TCR + IL-2: n = 7 per group; TCR + IL-33: n = 4 per group) or ST2⁺ cells (k) (TCR and TCR + IL-33: n = 4 per group; TCR + IL-2: n = 3 per group) gated on Treg cells was shown. l, m Percentage of GATA3⁺ST2⁺ (double positive, DP) cells among total ST2⁺ Treg cells (l) or GATA3⁺ Treg cells (m) was shown. (TCR and TCR + IL-33: n = 3 per group; TCR + IL-2: n = 2 per group). n, o Splenic Treg cells sorted from Ctrl (Foxp3^Cre-YFPSetd2^f/+) or KO (Foxp3^Cre-YFPSetd2^f/f) mice were cultured with α-CD3, α-CD28, IL-2 and IL-33, and were infected with retrovirus expressing Thy1.1, Thy1.1-ST2, or Thy1.1-GATA3. The percentage of GATA3⁺ cells (n) or ST2⁺ cells (o) gated on Thy1.1⁺ Treg cells (Thy1.1⁺Foxp3⁺) was analyzed by flow cytometry. (Ctrl n = 3; KO n = 5). p LI LPLs were isolated from mice of indicated genotypes (n = 5, 9, 7, 7, respectively). The percentage of GATA3⁺ST2⁺ cells gated on Helios⁺ Treg cells (CD3⁺CD4⁺Foxp3⁺Helios⁺) was analyzed by flow cytometry and normalized to Il33^+/–Foxp3^CreSetd2^f/+ group as 1 based on littermates. b–d, f–h, j–p Data were means ± SEM. a–h Data were pooled from 3–7 mice per time point for each genotype. i–p Representative of 2–6 independent experiments. Source data are provided as a Source Data file.