Fig. 4. Amino acid sequence alignment of WNT2b proteins and expression of zebrafish wnt2b in the zebrafish embryo. (A) Alignment of zebrafish (Danio rerio, Dr), chick (Gallus gallus, Gg), mouse (Mus musculus, Mm), and human (Homo sapiens, Hs) deduced WNT2b amino acid sequences, in addition to that of human WNT2. Identical amino acids are indicated by dots, and gaps by dashes. Asterisks indicate conserved cysteine residues among the WNT family members. The zebrafish Wnt2b sequence shows homology to that of chick (71.4%), mouse (61.2%), and human WNT2b (61.0%) and human WNT2 (63.0%). The presence of a 17 amino acid stretch just after the initiation methionine residue, which is characteristic of WNT2b but not WNT2, indicates that our clone encodes zebrafish wnt2b. The GenBank accession number for zebrafish wnt2b is AF544026. (B and C) Whole-mount in situ hybridization of 22 hpf zebrafish embryos showing wnt2b expression from a dorsal view (B) and a lateral view (C). Expression can be detected in the tissue medial to the LPM at the somite level where the pectoral fin buds will form (arrowheads) and the developing eye. Anterior to the left. (D) Cross section of 22 hpf zebrafish embryo at the pectoral fin bud forming level. wnt2b is expressed ventral to the somites and medial to the pectoral fin-budding region of the LPM. nt, neural tube; s, somite; nc, notochord.

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