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. 2014 Mar 23;5(5):336–343. doi: 10.7150/jca.7202

Figure 3.

Figure 3

N-23 suppresses TGF-β signaling by inhibiting TGF-β-dependent recruitment of Smad3 to the PAI-1 promoter. (A) N-23 does not affect pSmad protein levels. MCF-7 cells were treated with or without TGF-β (1 ng/mL), E2 (10-8 M), or N-23 (10-6 M). Western blotting was used to examine the levels of pSmad2, pSmad3, Smad2, Smad3, and β-Actin. (B) N-23 inhibits TGF-β-dependent recruitment of Smad3 to the PAI-1 promoter. MCF-7 cells were cultured in the presence or absence of TGF-β (1 ng/mL), E2 (10-8 M), or N-23 (10-6 M). A ChIP assay was performed with control IgG or anti-Smad3 antibodies. Immunoprecipitated DNA was examined using real-time RT-PCR and primers specific for the PAI-1 promoter. Samples were normalized to the amount of input DNA. ** indicates p < 0.01 and * indicates p < 0.05.