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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 3. The canonical Myb binding site in the CGRE is essential for the strong enhancer activity of the CGRE for IL-13 expression by c-Myb and GATA-3.

Figure 3

(A) Left schema shows CGRE and IL-13 promoter locus and Myb and GATA binding sites in CGRE. Dual-luciferase reporter assays were performed in 293T cells 24 hours after cotransfection with IL-13 promoter (IL13P), wild-type CGRE-IL13 promoter (CGRE-IL13P) and CGRE-IL13 promoter containing a mutated canonical Myb binding site within the CGRE (mut Myb). (B) Upper schema shows GATA binding sites and the generated mutation sites in CGRE. Reporter assays carried out in 293T cells 24 hours after co-transfection with CGRE-IL13P, CGRE-IL13 promoter containing a mutated canonical Myb binding site within the CGRE (mut Myb), CGRE-IL13P containing a mutated (shown with X in upper schema) high affinity GATA-3 binding site (mut GATA#3), or CGRE-IL13P containing four mutated GATA binding sites (mut GATA#1-#4). (C) Dual-luciferase reporter assays were performed with Jurkat cells with CGRE-IL13P, or CGRE-IL13P containing a mutated Myb binding site (CGRE-IL13P mut Myb). Cells were stimulated 24 hours post-transfection with PMA (200ng/mL) and ionomycin (300ng/mL) for 24 hours.