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Figure 4


Fig. 4. LRP6-initiated dephospho-β-catenin is transcriptionally more active than downstream-initiated dephospho-β-catenin. Cadherin-deficient SK-BR-3 breast carcinoma cells were transiently transfected with 25 or 50 ng plasmid encoding wild-type β-catenin or 160 ng of a plasmid encoding {Delta}N-LRP6 and analyzed by immunolocalization (A,B), TCF transcriptional activity (C) and western blotting (D,E). (A,B) Immunolocalization of dephospho-β-catenin in cells exogenously expressing β-catenin or {Delta}N-LRP6, identified by coexpression of mRFP-tagged histone H2B. (C) TCF-dependent transcriptional activity in SK-BR3 cells transfected with indicated plasmids 24 hours after transfection. TOP, TCF-reporter luciferase activity; FOP, mutated TCF-reporter activity. Values were normalized to a transfection control (constitutive Renilla luciferase reporter). (D,E) Western blot analysis of cells shown in A-C, detecting total (D) or dephospho-β-catenin (E) levels. 4x, fourfold amount loaded; 0.5x, one half amount loaded. Asterisk indicates a cross-reacting protein.





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