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. Author manuscript; available in PMC: 2012 Dec 1.
Published in final edited form as: J Immunol. 2011 Oct 28;187(11):5974–5982. doi: 10.4049/jimmunol.1100550

Figure 7. c-Myb silencing decreases methylation of histone H3K4 and acetylation of histone H3K9 at the IL-13 locus in stimulated primary human CD4+ effector/memory T cells.

Figure 7

Representative results of ChIP assays with a series of primer pairs covering the IL-13 and IL-4 gene locus in primary human CD4+ effector/memory T cells (A), and naïve T cells (B) stimulated under Th2 promoting conditions, expressed with c-myb shRNA or control scramble shRNA using the lentivirus systems. The human CD4+CD45RA+ T-cells (naïve) were cultured for 7 days under Th2 promoting conditions and CD4+CD45RA- T-cells (effector/memory) were cultured for 4 days with stimulation prior to ChIP assay. ChIP assays were carried out with antibodies directed against di- and tri- methylated HistoneH3K4 (upper graph in each panel), acetyl H3K9 (middle graph in each panel), and di-methyl H3K9 (bottom graph in each panel) and analyzed by QRT-PCR. Line graphs show the relative specific antibody mediated enrichment in comparison to control mouse, or rabbit IgG. The schematic below the base line of each top panel illustrates ~35 kb of the Th2 gene locus including the IL-13 and IL-4 genes. Four regulatory elements are shown with blank rectangles.