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. 2018 Jul 2;215(7):1853–1868. doi: 10.1084/jem.20170779

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© 2018 Tanaka et al.

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Figure 7. — Cooperative regulation of IL-17 and IL-10 by Trim33, Smad2, and ROR-γ. (A) Trim33 binding was assessed by ChIP-PCR analysis. PCR amplification was conducted with input DNA and I.P. DNA prepared from chromatin precipitated with anti-Trim33 antibodies, with C57BL/6 CD4⁺ T cells stimulated by anti-CD3 and anti-CD28 antibodies under indicated conditions. Primer for the H19 ICR locus was used as a negative control. Data are representative of three independent experiments. (B) Trim33 ChIP was performed with WT and Smad2 KO CD4⁺ T cells cultured under Th17 skewing conditions. (C) Physical association of Trim33 with Smad2 and ROR-γ in Th17 cells. Lysates of Th17 cells were subjected to immunoprecipitation with anti-Trim33 antibody or isotype Ig, followed by immunoblot analysis with anti-Trim33, Smad2, and ROR-γ antibodies. Data are representative of four independent experiments. Error bars show means ± SD. *, P < 0.05; **, P < 0.01.