Fig. 1. Expression and regulation of Dec2.

(a) Dec2 mRNA expression in T_H1, T_H2, T_H-17 or iT_reg cells was measured by quantitative RT-PCR. Dec2 expression in naïve CD4⁺ T cells was set as 1. (b) Kinetic expression of Dec2, Gata3 and Tbx21 mRNA in T_H1 or T_H2 cells. (c) Lung-associated lymph node cells from 4-get mice with asthma were restimulated with OVA for 48 hr.and IL-4-GFP⁺ and GFP⁻ cells were sorted on a CD4⁺CD44^hi gate and mRNA expression was examined by real-time RT-PCR. (d, e) Naïve CD4⁺ T cells were activated with the indicated plate-bound antibodies (d) or with plate-bound anti-CD3 and anti-CD28 in the presence or absence of IL-25 (e) and Dec2 mRNA expression was analyzed daily. (f) Dec2 and Il4 mRNA expression in naïve CD4⁺ T cells from Il4^−/− or Il4^+/+ mice infected with retrovirus expressing GATA3-IRES-GFP (GATA3) or GFP only (Vec).(g) Dec2 and Il13 mRNA expression in naïve CD4⁺ T cells from Gata3^fl/fl or Gata3^+/+ mice polarized under T_H2 conditions, infected with the bicistronic retrovirus hCre-IRES-GFP and further cultured under T_H2 conditions for 1 or 2 weeks. (h) Dec2 and Il13 mRNA expression in naïve CD4⁺ T cells from Gata3^fl/fl mice activated under T_H2 conditions and infected with a Cre-encoding virus or a control virus. (f–h) GFP⁺ fractions were sorted and analyzed. Data were normalized to Actb (a–f, h) or Cd4 (g) and show mean and s.d. (b–h) The sample with lowest detectable expression was set as 1. All experiments were repeated 2–3 times with consistent results.