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. 2015 Apr 1;128(7):1408–1421. doi: 10.1242/jcs.164467

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© 2015. Published by The Company of Biologists Ltd

This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/3.0), which permits unrestricted use, distribution and reproduction in any medium provided that the original work is properly attributed.

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Fig. 2. — Sulf1 inhibits the ability of Wnt8a to activate canonical Wnt signalling. (A) A graph showing the response of the Topflash reporter in whole embryos and (B) in animal caps injected with the mRNAs indicated. Results are mean±s.e.m. (C) An RNase protection analysis shows the expression of Chordin and Siamois in gastrula stage 10 embryos and in animal caps injected with mRNAs indicated. The expression of ODC serves as a loading control, and hybridisation to tRNA controls for specificity. (D) Western blotting for β-catenin shows protein levels in animal caps injected with the mRNAs indicated. Antibodies against MCM and GAPDH serve as loading controls. (E) Xenopus laevis embryos were microinjected with mRNA coding for β-catenin delta-N (labelled as β-catenin, 150 pg) or chordin (150 pg), alone or together with Sulf1 (1 ng) into a single ventral blastomere at the four-cell stage. The number of embryos with duplicated axes was counted at NF stage 20. (F) Immunoprecipitation of epitope-tagged proteins expressed in animal caps injected with the mRNAs indicated. The top panel shows protein immunoprecipitated with an antibody against HA (Wnt8a is tagged with HA) and immunoblotted with an antibodies against Myc (LRP6 is tagged with Myc). The bottom two panels are the protein lysates prior to immunoprecipitation.