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. 2016 May 31;113(24):E3451–E3460. doi: 10.1073/pnas.1506113113

Fig. 6.

Fig. 6.

T3 represses stimulation of endogenous TGF-β target genes and inhibits in vivo occupancy of the promoters by SMADs. (A) Level of transcripts for the Smad7 and p15 genes were determined by quantitative PCR in GH4C1 cells treated with T3 for 36 h and TGF-β for the last 24 h. The early induction of Id1 mRNA was analyzed after 1 h treatment with TGF-β. Data are mean ± SD. Significance of Bonferroni post hoc test (n = 3) is indicated. (B) ChIP assays were performed with the indicated fragments of the TGF-β target promoters that contain SBEs. Location of the primers used is shown with arrows, and the SBEs are depicted as black boxes. Cells were treated for 1 h with T3 and/or TGF-β and analyzed by ChIP with noninmmune IgGs, SMAD2/3, SMAD4, TRβ, and acetylated histone H4 antibodies. One representative experiment of two is shown. The input for each assay is also shown.