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Fig. 7. Sox17β requires β-catenin to robustly activate target gene transcription in animal caps. (A) Embryos were injected at the two-cell stage with the indicated combinations of: Sox17β mRNA (200 pg), RNA encoding a N-terminal deleted form of stabilized β-catenin (δN-β-catenin, 100 pg) (Yost et al., 1996), or an antisense β-catenin morpholino oligos (βcat-MO; 20 ng). At blastula stage, animal cap tissue was explanted and cultured for 3-4 hours until gastrula stage, when it was assayed by real-time RT-PCR for the expression of Sox17 target genes. The histograms show the relative expression levels normalized to the loading control, ODC. Plakoglobin (Plako) is control gene that is neither a Sox17 nor β-catenin target. (B) A proportion of each sample from the same experiment was assayed by immunoblotting with anti-β-catenin, anti-Sox17β or anti-tubulin antibodies. Injected δN-β-catenin protein has a higher molecular weight than endogenous β-catenin because of the presence of an epitope tag. Tubulin is a loading control. 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. Experiment + Assay Source Phenotypes and Disease Xla Wt + ctnnb1del + sox17b.1 + ctnnb1 MO + animal cap explant + gastrula stage (RT-PCR) fig.7 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 Xla Wt + sox17b.1 + animal cap explant + gastrula stage (RT-PCR) fig.7 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 Larger Image Printer Friendly View

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