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Fig. 6. In situ hybridization analysis for putative Pitx3 target genes Obscnl and Baz2b. Visual comparisons of gene expression patterns between control-morpholino (Cmo) and Pitx3-morpho- lino (Pmo) right side-injected embryos. A: Obscnl shows a loss of expression in the branchial arches (black arrow), otic vesicle, and retina when treated at stage 27 with Pmo (A0) versus Cmo (A). B: Baz2b is substantially reduced in response to Pmo (B0) in the retinal layer of the optic protuberance (white arrow), as well as in the pronephros and in the anterior region of the dorsal axis, when compared with Cmo (B). |
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Fig. 7. Characterization of a novel transcript, Baz2b, in X. laevis. A: Protein alignment showing amino acid similarities between Baz2B homologs across organisms. B: Temporal expression of Baz2b throughout embryonic stages of development show expression as a maternal transcript in the egg E and throughout development to tailbud stage, with slight reductions in transcript level at stages 10 and 19. B Confirmation of microarray predictions by means of reverse tran- scriptase-polymerase chain reaction (RT-PCR) show abolished expression at stage 27 in response to Pitx3-morpholino (Pmo) when compared with control-morpholino (Cmo) and wild- type (WT) embryos. C: Baz2b expression at stages 21 (C), 31 (D), and 35 (E) is concentrated in the developing eye, as well as the branchial arches and otic vesicle. Dark expression is seen in the pronephros, persisting in the tubules (E). F: A schematic diagram of Baz2b protein depict- ing various domains characteristic of Baz2B: methyl-CpG binding domain (MBD), DNA binding domain (DDT), zinc finger domain (Z), adjacent to the bromodomain (BR). GenBank accession numbers used to generate phylogenetic tree (A) are as follows: xBaz2b BQ400337 (X. laevis), mBaz2b BC150814 (mouse), rBaz2b NM_001108260 (rat), hBAZ2B NM_013450 (human), cBaz2b NM_204677 (chick), xtBaz2b BC166361 (X. tropicalis). |
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bazb2 (bromodomain adjacent to zinc finger domain, 2B) gene expression in Xenopus laevis embryo, assayed via in situ hybridization, NF stage 31, lateral view, anterior left, dorsal up. |
