FIGURE 2.

The transactivational activity for the Il5 promoter is impaired by methylated Gata3. A and B, reporter assays with the Il5 promoter and the Il13 promoter were performed using the M12 cell line that expresses no Gata3 family proteins. WT or mutant Gata3 cDNAs were introduced, and the luciferase activity was measured. The data indicate the mean results of three independent experiments with standard deviations. Mock, mock-transfected. **, p < 0.01 by Student's t test. C, 293T cells were transfected with an expression plasmid encoding FLAG-tagged Gata3 WT or one of the mutants, and total extracts were subjected a pulldown assay using Il5 promoter oligonucleotides (Oligo) as described under “Experimental Procedures.” The results of immunoblotting of the total lysates are also shown. D, naive CD4 T cells were stimulated under Th1 conditions and then infected with a retrovirus vector carrying WT or mutant Gata3 cDNAs. Four days later, the status of H3-K9 Ac and H3-K4 3Me in the Th2 cytokines and the Ifng promoters was determined by a ChIP assay with a qPCR analysis. The relative intensities (/input) are shown with standard deviations.