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. 2015 Apr 27;290(23):14717–14728. doi: 10.1074/jbc.M114.630368

FIGURE 6.

FIGURE 6.

Direct binding of SMAD4 to the CCL20 SMAD binding element (SBE2). A, EMSA with a biotinylated probe containing SBE2 of the human CCL20 promoter and recombinant SMAD4 (lane 1) in the presence of WT (lane 2) or SBE2 mutant oligonucleotides (lane 3) or anti-SMAD4 (lane 4). The positions of three bands and a fourth supershifted band with anti-SMAD4 are shown. B, EMSA using nuclear extracts of human lung fibroblasts treated with vehicle control (lanes 1–3) or recombinant IL-1β (lanes 3–6) in the presence of WT (lanes 2 and 5) or SBE2 mutant oligonucleotides (lanes 3 and 6). The positions of the three lower bands are similar to those seen in A. C, EMSA using nuclear extracts of human lung fibroblasts treated with recombinant IL-1β with no antibody (lane 1), control IgG (lane 2), anti-SMAD3 (lane 3), or anti-SMAD4 (lane 4). Supershifts with anti-SMAD3 (lane 4) or competition with anti-SMAD4 (lane 4) are shown. Mut, mutant; oligo, oligonucleotide.