c-Myb silencing decreases the methylation of histone H3K4 and the acetylation of histone H3K9 at GATA-3 locus in Jurkat cells, and primary human CD4⁺ naive and effector/memory cells with the stimulation under Th2-promoting conditions. Representative result of ChIP assays carried out with a series of primer pairs covering the GATA-3 gene locus in Jurkat cells (A), primary human CD4⁺ CD45RA⁺ naive T cells (B), and CD4⁺CD45RA⁻ effector/memory T cells (C) infected with lentivirus expressing either c-Myb or control (scrambled sequence) shRNA. Cells were stimulated under Th2-promoting conditions for 7 days and 4 days, respectively, before conducting the ChIP assays. The ChIP assays were carried out with antibodies directed against dimethylated and trimethylated histone H3K4 (top graph in each panel), acetyl H3K9 (middle graph in each panel), and dimethyl H3K9 (bottom graph in each panel) and analyzed by quantitative RT-PCR. The graphs show the relative specific antibody-mediated enrichment compared with control mouse or rabbit IgG. Each point represents the mean ± SD of 3 replicate determinations. In each top panel of the figure, the line drawing immediately underneath the baseline of each graph illustrates (A) approximately 27 kbp or (B-C) approximately 20 kbp of the GATA-3 gene locus. The previously validated, highly conserved c-Myb binding site is situated between primer pairs 4 and 5. Two independent experiments were performed with similar results. *P < .05. **P < .01.