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. 2011 Oct 24;208(11):2321–2333. doi: 10.1084/jem.20110462

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© 2011 Ruan et al.

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Figure 8. — Rel controls Rorg promoter activities in primary Th17 cells. (A and B) Purified naive CD4⁺ T cells from WT (n = 3) mice were cultured under Th17 differentiation conditions, as described in Materials and methods. At indicated times, cells were fixed, and ChIP was performed using anti-p65 antibody. Re-ChIP was performed with anti–c-Rel using DNA precipitated by anti-p65. (C and D) Purified naive CD4⁺ T cells from 6-wk-old Rel^−/− mice (n = 3) were cultured under Th17 differentiation conditions, as described in Materials and methods. After 17 h, cells were fixed, and ChIP was performed using anti-p65 antibody. (E–G) Naive CD4⁺ T cells from WT (E–G; n = 3) and Rel^−/− mice (E; n = 3) were transiently transfected with WT (E–G) and Rel sites mutated (E) murine Rorgt promoter luciferase constructs using Amaxa Nucleofector reagent. An expression vector for full-length c-Rel was also used to co-transfect cells in E. Cells were cultured under Th17 differentiation condition for 24 h, and the luciferase activities measured. To normalize the transfection efficiency across samples, the Renilla luciferase expression vector pRLTK was included as an internal control. Error bars indicate the SEM. Data are representative of two independent experiments.