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Fig. 6. Dpr immunoprecipitation, localization and activation of Wnt/ß-catenin target genes. (A) Dpr1-myc and (B) Dpr2-myc immunoprecipitate with Dvl2-HA from Xenopus lysates. (C) Dpr1-myc and (D) Dpr2-myc localization in Xenopus animal caps. Small arrow in D indicates nuclear staining. Large arrow indicates membrane staining. Scale bar: 20 µm in C,D. (E) Injection of 4 ng dpr1 RNA induces ß-catenin target genes, but 4 ng dpr2 RNA does not. (F) Dpr1 and Dpr2 synergize with suboptimal dose of zebrafish Dvl2. dpr1 and dpr2 low dose is 0.25 ng, medium dose is 0.5 ng and high dose is 1.0 ng. ß-galactosidase RNA dose is 1.0 ng. (G) Dpr1 and Dpr2 with internal control ß-gal in TnT reaction (upper). Dpr1-myc and Dpr2-myc western from Xenopus lysates with internal control GFP (lower). (H) Dpr1-Dvl2 synergy is inhibited by Axin. (I) Dpr1 and Dpr2 do not inhibit Wnt8 activation of ß-catenin target genes. (J) Dpr1 and Dpr2 synergize with Dvl2 in activating a ß-catenin-dependent luciferase reporter in HEK 293T cells. The experiment is representative of three independent transfections, with each data point being conducted in triplicate. Unless noted otherwise, 1 ng of dpr1-myc, dpr2-myc and ß-galactosidase RNA, and 0.4 ng dvl2 RNA were used. For I, 1 pg of wnt8 RNA, and 4 ng of dpr1 and dpr2 RNA were used. For J, 400 ng of Dpr1, Dpr2 or ß-galactosidase, and 20 ng of zebrafish Dvl2 were transfected. AU, arbitrary light units.





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