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. Author manuscript; available in PMC: 2016 Mar 14.
Published in final edited form as: Nat Immunol. 2015 Feb 2;16(3):286–295. doi: 10.1038/ni.3099

FIGURE 7. IL-1β reverses RA-mediated STAT5 binding in the Il17a-Il17f and Foxp3 gene loci.

FIGURE 7

(a,b) Chromatin immunoprecipitation (ChIP) analysis of STAT3 and STAT5 binding in purified CD4+CD8− thymocytes isolated from wild-type (WT) or Stat5fl/fl.Cd4-cre mice and activated with plate-bound anti-CD3 and soluble anti-CD28 under TH0 or TH17 conditions (IL-6 +TGF-β, with or without the indicated additions of RA and IL-1β. On day 4, recovered cells were stimulated with IL-6 (pSTAT3) or IL-6+IL-2 (pSTAT5) and processed for ChIP-PCR to quantitate binding of STAT3 or STAT5 to the indicated sites within the Il17a-f (a) and Foxp3 (d) gene loci. Unstimulated TH0 cells were used as a negative control. Results are relative to input DNA. Data are representative of two independent experiments with 3 replicates per experiment (means and s.e.m. in a, b). *P < 0.05 and **P < 0.01 (two-tailed unpaired t-test).