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. 2010 Sep 30;285(49):38023–38033. doi: 10.1074/jbc.M110.132209

FIGURE 2.

FIGURE 2.

Cross-talk between TGF-β and Wnt signalings to activate the TMEPAI gene. A, expression of TMEPAI mRNA in intestinal adenoma of ApcMin/+ mice. WT, intestinal mucosa of wild-type mice; ApcMin/+, intestinal polyps of ApcMin/+ mice. B, reduction of TGF-β-induced TMEPAI mRNA in TCF7L2 knocked down MCF10A1 cells. MCF10A1 cells carrying nontargeting shRNA (control shRNA) or a mixture of TCF7L2 shRNAs were stimulated with TGF-β for 2 h. Subsequently, the expression of TCF7L2 mRNA was checked using RT-PCR. Because TCF7L2 possesses alternative splicing forms, two PCR products could be seen using primers used here. C, activation of pGL3ti-850 by TCF7L2 or LEF1. HepG2 cells were transfected with a different amount of TCF7L2 or LEF1 together with pGL3ti-850 (upper panel). Simultaneously, the expression of TCF7L2 or LEF1 proteins in HepG2 cells was detected by Western blot (WB) analysis (lower panel). D, synergistic activation of pGL3ti-850 by the combination of ALK5ca with lithium chloride. 293T cells transfected with pGL3ti-850 together with or without ALK5ca were cultured in the presence or absence of 20 mm lithium chloride. E, inhibition of TGF-β-induced pGL3ti-850 activity by TCF7L2Δ(1–30). HepG2 cells transfected with TCF7L2Δ(1–30) were stimulated with TGF-β. F, synergistic activation of pGL-3ti-850. HepG2 cells transfected with Smad3, TCF7L2, β-catenin, or their combinations were stimulated with TGF-β. G, requirement of DNA binding ability in Smad proteins for synergistic activation of pGL3ti-850. Smad3-deficient MEF cells transfected with Smad2, Smad2Δexon3, Smad3, and/or TCF7L2 were stimulated with TGF-β.