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. Author manuscript; available in PMC: 2020 Oct 1.
Published in final edited form as: Mol Immunol. 2019 Aug 6;114:260–268. doi: 10.1016/j.molimm.2019.07.029

Figure 5. SKI requires SMAD4 to modulate Rorc gene transcription.

Figure 5.

(a) CD4+ T cells isolated from wild-type (WT) or (b) Cd4-cre;Smad4fl/fl (SMAD4 KO) mice were cultured with IL-21 and TGF-β, and then transduced with control retrovirus (RV) or SKI-expressing retrovirus (RV SKI). IL-17A+ cells were analyzed by flow cytometry. Representative results and statistical analysis are shown. (c) CD4+ T cells isolated from wild-type (WT) mice were cultured with IL-21 and TGF-β, and then transduced with control retrovirus (RV), SKI-expressing retrovirus (RV SKI), or SKI and RORγt co-expressing retrovirus (RV SKI+RORγt). IL-17A+ cells were analyzed by flow cytometry. Representative results and statistical analysis are shown. (d) CD4+ T cells isolated from wild-type (WT) mice were cultured under indicated conditions, cells were harvested after 3 days. ChIP assay was performed with control IgG antibody or SKI antibody. The relative enrichment of SKI binding to the Rorc locus was determined. (e) CD4+ T cells isolated from wild-type (WT) and Cd4-cre;Smad4fl/fl (SMAD4 KO) mice were cultured with IL-21 and TGF-β receptor I inhibitor, cells were harvested after 3 days. ChIP assay was performed with control IgG antibody or SKI antibody. The relative enrichment of SKI binding to the Rorc locus was determined.