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. 1999 Oct 18;147(2):367–374. doi: 10.1083/jcb.147.2.367

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© 1999 The Rockefeller University Press

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 4.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/4.0/).

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Target gene induction and β-catenin dose responsiveness in the cell-free assay system. A, Increasing amounts of β-catenin protein (as indicated) were added to 20 crushed embryos under standard cell-free assay conditions. RT-PCR (upper panel) analysis of Siamois expression revealed a dose-dependent induction. Western blot analysis of total lysate and ConA-Sepharose– treated (glycoprotein depleted) lysate was carried out using antibodies to the NH₂-terminal region of β-catenin (lower two panels). Endogenous and free β-catenin show different electrophoretic mobility, due to the presence of a 6× His tag on the exogenous protein. Incubation of extracts with ConA-Sepharose effectively depleted C-cadherin below detectable levels (data not shown). B, In addition to Siamois, addition of β-catenin (1.5 μg/20 embryo condition) induces expression of XTwin and Xnr3 mRNA, as demonstrated by RT-PCR.