Decreased production of IFN-γ by NFAT1^−/− IL-4^−/− T cells cannot be explained by overexpression of Maf or GATA-3.

CD4⁺ T cells from NFAT1^+/+IL-4^−/− and NFAT1^−/− IL-4^−/−mice were cultured for 4 days with anti-CD3 and IL-2, either under neutral conditions (lanes 1 and 2) or in the presence of 1000 U/mL IL-4 to induce Th2 differentiation (lanes 3 and 4). After harvesting the cells, total RNA was extracted, and Northern blot analysis was performed as described in “Materials and methods.” Ethidium bromide staining of the 28S bands confirmed equal loading of RNAs isolated from NFAT1^−/− IL-4^−/− and NFAT1^+/+IL-4^−/− cells (data not shown). Results of 1 of 2 identical experiments are shown.