Figure 4. Enforced expression of Bach2 restores the phenotypes of the Menin KO effector CD4 T cells.

(a) The results of the quantitative RT-PCR analysis of mRNA encoding Bach2 in the WT and Menin KO naive CD4 T (right) and T_H (left). The results are presented relative to the mRNA expression of Hprt with the s.d. (b) The results of the immunoblot analysis of Bach2 in the nuclear fractions of the WT and Menin KO T_H cells. (c) The results of the intracellular FACS analysis of IL-4/IFN-γ and IL-2/IL-6 in the Menin KO T_H cells transduced with an empty vector (Mock) or with Bach2-expressing retroviral vector (Bach2). The naive CD4 T cells cultured under IL-2-conditions were infected with retroviral vectors and cultured for 3 days in the presence of IL-2. (d) The mock- or Bach2-transduced cells in c were purified using an AutoMACS device and restimulated with immobilized anti-TCR-β for 16 h. The amounts of cytokines in the supernatants were determined using ELISA. (e) The amounts of pro-inflammatory chemokines in the supernatants in d were determined using ELISA. (f) The results of the quantitative RT-PCR analysis of the Gzm’s and Prf1 of cells in d. The results are presented relative to the mRNA expression of Hprt with the s.d. All experiments were performed with T_H cells cultured under IL-2 conditions for 5 days. Three independent experiments (a–f) were performed. **P<0.01 (Student’s t-test).