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Fig. 4 Phylogenetic analysis of zebrafish NCX isoforms and expression patterns of ncx3 and ncx4b. (A) The sequences of zebrafish ncx1h, ncx1n, ncx3 and ncx4a were derived from RACE and RT-PCR. Full-length zebrafish ncx2 and ncx4b sequences were obtained from the Ensembl Genome Browser. The amino acid sequences of human, mouse and fugu NCX isoforms are the same as those used by Marshall et al. (Marshall et al., 2005). Sequence alignments and phylogenetic analyses were performed using ClustalX. The unit of the scale bar represents 10% amino acid substitution per site. (B) ncx3 is expressed predominantly in the somites in 1-day-old zebrafish embryos. (C) ncx4b expression is restricted in the hypaxial muscle after 3 days of development.