Figure 4. IL-2 suppression and IFN-γ induction by T-bet in Th cells. Spleens and lymph node tissues were collected from WT and DTg/KO mice and CD4⁺ Th cells were isolated. Isolated Th cells were stimulated with anti-CD3 (2 µg/mL) in the either presence (+) or absence (-) of doxycycline (0.5 µg/mL) for 3 days. (A) Cell pellets were harvested and subjected to protein analysis by immunoblotting with Ab against T-bet or actin. (B) Cell supernatants were harvested for IL-2 and IFN-γ analysis by ELISA. Five independent experiments were performed and data are expressed as the average ±SD for 5 mice per group. ^*p<0.05; ^***p<0.0005.