(A) Naïve CD4+ T cells were activated for 48 hours in the absence of skewing cytokines before being transduced with control or STAT3C-expressing retrovirus. After 5 days in culture, cells were sorted and re-stimulated with anti-CD3 (4 μg/ml) for 24 hours before cell-free supernatant was tested for cytokines using ELISA (mean ± s.d.).
(B) Naïve CD4+ T cells were activated for 48 hours under Th1 conditions (5 ng/ml IL–12 + 10 μg/ml anti-IL-4) before being transduced with control, STAT6VT-expressing, STAT3-expressing or both retroviruses. After 5 days in culture, cells were sorted and re-stimulated with anti-CD3 (4 μg/ml) for 24 hours before cell-free supernatant was tested for cytokines using ELISA (mean ± s.d.).
(C) CD4+ cells were isolated from WT, STAT6VT and STAT6VT-Stat3Cd4−/ − mice. Cells were then re-stimulated with anti-CD3 for 24 hours. Cell-free supernatant was collected and tested for various cytokines using ELISA. Data are representative of 2 experiments (average of 2–6 mice ± s.d.). Students t test was performed to calculate p values.