Fig. 2. Gene product of Cs-FoxD. (A) Nucleotide and deduced amino acid sequence of Cs-FoxD clone. The 1915 bp insert includes a single open reading frame that encodes a polypeptide of 506 amino acids. The termination codon is shown by an asterisk. The predicted forkhead domain is shown by bold letters. The nucleotide sequence of the 5' region used to prepare morpholino oligonucleotides is underlined. (B) Comparison of amino acid sequences of the forkhead domains of Cs-FoxA5, Cs-FoxD and zebrafish z-FoxD1. (C) Molecular phylogenetic analysis of relationships of Cs-FoxD with Fox gene members. The tree was constructed by comparison of the amino acid sequences of the forkhead domains using the neighbor-joining method. Human FoxC1 and FoxC2 were used as the outgroup. Branch lengths are proportional to the evolutionary distance corrected for multiple substitutions with the scale denoting 0.01 amino acid substitutions per site. The numbers indicate the relative robustness of each node as assessed by bootstrap analysis (100 replications). This analysis strongly suggests that Cs-FoxD is a member of the FoxD subclass.