Xenbase Image ID: 156717

Figure 3. XSox17β Represses Wnt/β-catenin Stimulated Transcription and Activates Endodermal Genes (A and B) Human 293T cells were cotransfected with OT reporter construct containing TCF consensus binding sites (Korinek et al. 1997) and expression plasmids encoding (A) S37A-β-catenin (0.2 μg), S37A-β-catenin (0.2 μg) + XSox17β (2 μg and 4 μg) and (B) Wnt1 (2 μg), Wnt1 (2 μg) + XSox17β (2 μg and 4 μg). XSox17β inhibits β-catenin and Wnt1 activation of the OT reporter. (C) Schematic of XSox17β:GAL4 DNA-binding domain fusion constructs and the 5x GAL DNA binding site–luciferase construct. (D) Human 293T cells cotransfected with 5x GAL reporter (0.5 μg) and 1 μg of the indicated fusion construct, with (gray bars) or without (black bars) 1 μg of S37A-β-catenin. The C-terminal region of XSox17β has transactivation capacity. Transfection experiments were repeated two to four times, and a representative experiment is shown. (E) Embryos were injected with mRNA encoding XSox17β (125 pg, 250 pg, 500 pg, 1000 pg), pt-β-catenin (30 pg), and pt-β-catenin (30 pg), + XSox17β (125 pg, 250 pg, 500 pg, 1000 pg). Animal caps dissected at blastula stage were cultured until gastrula stage and (F) analyzed by RT-PCR for expression of the Wnt/β-catenin responsive genes Siamois and Xnr3, the endodermal gene Endodermin, and Ef-1α as a loading control. Image published in: Zorn AM et al. (1999) Copyright © 1999. Image reproduced with permission of the Publisher, Elsevier B. V. Experiment + Assay Source Phenotypes and Disease Xla Wt + ctnnb1tm1 + animal cap explant + gastrula stage (RT-PCR) fig.3.f Expression Phenotype increased amount nodal3.1.L expression in animal cap increased amount sia1.L expression in animal cap Xla Wt + ctnnb1tm1 + sox17b.2 + animal cap explant + gastrula stage (RT-PCR) fig.3.f Expression Phenotype increased amount a2m.L expression in animal cap Xla Wt + sox17b.2 + animal cap explant + gastrula stage (RT-PCR) fig.3.f Expression Phenotype increased amount a2m.L expression in animal cap Larger Image Printer Friendly View

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