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Fig. S1 Functional overexpression of recombinant Wnt3. The recombinant Wnt3 zebrafish fusion protein with EGFP at the C-terminal (Wnt3EGFP) is functional as overexpression in zebrafish produces a posteriorized brain phenotype typical of overt Wnt activity. (A) Bright field image of a malformed embryo with a posteriorized brain upon overt Wnt3EGFP expression under the ubiquitous CMV promoter. Injection of the Wnt3EGFP expression construct at blastula stage resulted in the deformed phenotype. (B) Tg(memKR15-16) expressed membrane localized KillerRed in the midbrain hindbrain boundary (mhb) was used to characterize the deformed brain phenotype. (C) The posteriorized brain phenotype is accompanied by the loss of the midbrain hindbrain boundary observed in Tg(memKR15-16) upon overt Wnt3EGFP expression.