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Fig. 5. Wise requires components of the Wnt pathway for en2 induction and activates the canonical Wnt pathway. (A,B) RT-PCR of noggin-injected animal caps assayed for en2 induction. Induction of en2 is attenuated or blocked by dominant-negative (dn)-Wnt8 ({Delta}Wnt8), dn-LRP6 ({Delta}LRP6), dn-Dishevelled [{Delta}Dsh (dd1)], GSK3-ß (GSK3), dn-Lef1 (LEF{Delta}N) (A) and dn-Frizzled8 ({Delta}Fz8) (B). (C) Wise does not interfere with the ability of Wint8 to induce Krox20 in noggin-injected animal caps. Wnt8-induced Krox20 expression is blocked by {Delta}Wnt8, but not by Wise. (D) Long-exposure of RT-PCR result showing that Wise-injected animal caps express siamois and Xnr3, although very weakly in comparison with the induction by Wnt8. (E-G) Staining for subcellular localisation of endogenous ß-catenin detected immunocytochemically in Xenopus animal caps after RNA injection of (E) Tcf3, (F) Wnt8+Tcf3 and (G) Wise+Tcf3. Both Wnt8 and Wise promote nuclear accumulation of ß-catenin.





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