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Fig. 1. Phylogenetic analysis and early expression pattern of XBarhl2. (A)
Phylogenetic tree based on identity in 60 amino acid homeobox region between
the Barhl proteins in human, mouse, rat, Xenopus, C. elegans and
Drosophila and the Barx proteins of human and chicken
(Barlow and Francis-West, 1997;
Edelman et al., 2000;
Hjalt and Murray, 1999). Mouse
Barhl2 (Mo et al., 2004),
mouse Barhl1 (Bulfone et al.,
2000; Li et al.,
2002) and XBarhl2 (Patterson
et al., 2000; Poggi et al.,
2004) sequences were determined by us. Human Barhl2 (GenBank
AL355867) and human Barhl1 (GenBank AL354735) sequences were predicted from
their genomic DNA sequences; rat Barhl2 (MBH1)
(Saba et al., 2003;
Saito et al., 1998). The tree
was determined using MegAlign from the Lasergene Navigator, licensed by
DNAStar. d, Drosophila melanogaster; m, Mus musculus; r,
Rattus norvegicus; x, Xenopus laevis; d-rer, Dario
rerio; ce, Caenorhabditis elegans; h, Homo sapiens. (B)
Expression of Xbarhl2 as determined by RT-PCR analysis on embryos at
different embryonic stages and dissected parts of embryos. (a)
Xbarhl2 starts to be expressed at stage 10.5. XEF-1
was analyzed as a control. (b) At stage 10.5 Xbarhl2 is expressed in
the dorsal part of the embryo. (C,D) Whole-mount in situ hybridization of
embryos, using Xbarhl2 as a probe. (C) Dorsal view of stage 14
embryos showing expression of Xbarhl2 in the dorsal diencephalon. (D)
Same embryo as C rotated showing expression along the midline (arrow). A,
anterior; P, posterior. (E) RT-PCR analysis. (a) Drawing of a stage 14 embryo
showing dissected parts (Nieuwkoop and
Faber, 1967). (b) Xbarhl2 is expressed in the posterior
neural plate, posterior dorsal mesoderm and in the midline of stage 14 embryo.
Chordin and Ncam were analyzed as mesodermal and
neuroectodermal markers, respectively.
