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. 2009 Mar 15;23(6):681–693. doi: 10.1101/gad.1773109

Figure 4.

Figure 4.

SMRT is recruited to the Il12b promoter through TEL (A) TEL binds specifically to the Il12b promoter. ChIP analysis for binding of TEL, IRF2, and METS to the Il12b promoter was performed with untreated (gray bars) and LPS-treated (black bars) primary macrophages using TEL, IRF2, and METS antibodies or control IgG (*) P < 0.05 versus IgG. (B) The κB and Ets sites are necessary for TEL binding to the Il12b promoter. RAW cells were transfected with the indicated wild-type and mutant Il12b-luciferase reporter plasmids. ChIP assay was performed with antibody against TEL and control IgG. Immunoprecipitated DNA was analyzed by real-time PCR using primers designed to amplify a region between the luciferase and the proximal promoter region. (*) P < 0.01 versus wild-type. (C) TEL and SMRT are present in the same complex on Il12b promoter as shown by ReChIP assay done in primary macrophages. (*) P < 0.05 versus IgG. (D) TEL is the beacon for the recruitment of SMRT on Il12b promoter. Primary macrophages were transfected with siRNA control and siRNA against TEL. ChIP assays were performed using antibodies against SMRT (top panel, gray bars) or control IgG (black bars, top and bottom panels). (*) P < 0.05 versus IgG. Results are expressed as average of three independent experiments. Error bars represent standard deviations. (E) Preferential co-IP of HA-TEL by Flag-SMRT. (F) Preferential co-IP of c-Jun by Flag-NCoR.