Fig. 1. Structure of pnx. Pnx is a novel homeodomain-containing protein. (A) Amino acid sequence of Pnx. Pnx contains an Eh1 (Engrailed homology 1) repressor domain in its N-terminal region (underlined) and a homeodomain in its C-terminal region (shaded box). (B) Phylogenetic tree for comparison between Pnx, Nkx1/Sax-family homeodomain-containing proteins and Gbx. The phylogenetic tree was constructed using the ClustalW program provided by DDBJ and the full-length amino acid sequences of the proteins (except for chick Hox3, which has only a C-terminal partial sequence). Accession Numbers of the proteins used for the phylogenetic tree are AB067731 (zebrafish Pnx), AB067732 (zebrafish Sax1), NP033149 (mouse Sax1), JU0069 (chicken Hox3), A33976 (Drosophila NK-1), AAG14450 (Xenopus BarH1), XP123759 (mouse BarH-like1), XP069853 (human Gbx1), XP144233 (mouse Gbx1) and AAK83070 (zebrafish Gbx1). (C) Alignment of the Eh1 repressor domains of zebrafish Pnx, Goosecoid, Fez-like, Dharma/Bozozok, and Engrailed3, and mouse Sax1. The consensus sequence for the Eh1 domain is also shown. (D,E) Nuclear localization of Pnx protein. COS7 cells were transfected with an expression vector for Myc epitope-tagged Pnx. Myc-Pnx was immunostained with an anti-Myc antibody. Pnx was detected in the nuclei of the transfected cells (indicated by arrowheads) (D). (E) A DAPI image of D. DAPI marks nuclei.

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