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Fig. 2. Direct targets of Sox17β. (A) A schematic of the GR:Sox17β fusion protein consisting of the hormone-binding domain of the human glucocorticoid receptor fused to Sox17β. (B) Isolated blastula animal cap explants either injected with mRNA encoding GR:Sox17β (150 pg) or uninjected were each cultured in three conditions: 1×MBS, 1×MBS + 10–6 dexamethasone (DEX) or in 1×MBS with 10–6 dexamethasone + 10 μg/ml cycloheximide (CHX). Dexamethasone activates the GR:Sox17β fusion protein and cycloheximide blocks translation. Control animal caps were treated with 5 ng/ml human activin A either with or without 10 μg/ml cycloheximide. At gastrula (stage 11) the explants were assayed by RT-PCR. GR:Sox17β induced Hnf1β, Foxa1, Foxa2 and Sox17α transcription when translation was blocked and are therefore direct Sox17 targets. A repeat of this experiment assayed by real-time RT-PCR is presented in Figs S1-S3 at http://dev.biologists.org/supplemental. (C) Whole-mount in situ hybridization to bisected gastrula (stage 11) embryos with the indicated probes confirms that Sox17 target genes are expressed in the endoderm. Dorsal/anterior towards the left. Image published in: Sinner D et al. (2004) Copyright © 2004. Image reproduced with permission of the publisher and the copyright holder. This is an Open Access article distributed under the terms of the Creative Commons Attribution License. Gene Synonyms Species Stage(s) Tissue a2m.S A2Moo, Alpha2-macroglobulin, edd, endod, endodermin, LOC108695707, ovomacroglobulin X. laevis Throughout NF stage 11 deep endoderm foxa1.L fkh2, foxa1-a, foxa1-b, foxa1b, HNF-3 alpha, HNF3 alpha, HNF3-alpha, hnf3a, HNF3alpha, tcf3a, tFoxA1, xfkh2 X. laevis Throughout NF stage 11 deep endoderm gata4.L gata-4, gata4-a, gata4-b, xGATA-4, xgata4 X. laevis Throughout NF stage 11 deep endoderm Image source: Published Experiment + Assay Source Phenotypes and Disease Xla Wt + Activin + animal cap explant + NF11 (RT-PCR) fig.2 Expression Phenotype increased amount a2m.L expression in animal cap increased amount foxa1.L expression in animal cap increased amount foxa2.L expression in animal cap increased amount gata4.L expression in animal cap increased amount gata6.L expression in animal cap increased amount gsc.L expression in animal cap increased amount hnf1b.L expression in animal cap increased amount nodal.L expression in animal cap increased amount sox17a.L expression in animal cap Xla Wt + Activin + CHX + animal cap explant + NF11 (RT-PCR) fig.2 Expression Phenotype increased amount a2m.L expression in animal cap increased amount chrd.1.L expression in animal cap increased amount gata4.L expression in animal cap increased amount gata6.L expression in animal cap increased amount gsc.L expression in animal cap increased amount hao1.L expression in animal cap increased amount nodal.L expression in animal cap Xla Wt + sox17b-GR + CHX + DEX + animal cap explant + NF11 (RT-PCR) fig.2 Expression Phenotype increased amount a2m.L expression in animal cap increased amount foxa1.L expression in animal cap increased amount foxa2.L expression in animal cap increased amount hnf1b.L expression in animal cap increased amount nodal.L expression in animal cap increased amount sox17a.L expression in animal cap Xla Wt + sox17b-GR + DEX + animal cap explant + NF11 (RT-PCR) fig.2 Expression Phenotype increased amount a2m.L expression in animal cap increased amount foxa1.L expression in animal cap increased amount foxa2.L expression in animal cap increased amount hao1.L expression in animal cap increased amount hnf1b.L expression in animal cap increased amount sox17a.L expression in animal cap Larger Image Printer Friendly View

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