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Supplementary material Supplement Fig. 5 EnR-Lef1-GR and Tcf3∆C-GR inhibit canonical Wnt signaling as indicated by ventralization ofXenopusembryos. A Schematic representation of wild type and dominant negative GR-fusion constructs of Tcf3A and Lef1 employed for repression of canonical Wnt signaling. B Experimental set up: embryos were injected with the respective constructs in both dorsal blastomeres at the 4-cell stage, treated with Dexamethasone at the 16-cell stage, and ventralization was analyzed at early tailbud stages. C Graph summarizing the percentage of ventralized embryos in 3 independent experiments after injection of EnR-Lef1-GR. At low concentrations EnR-Lef1-GR overexpression did not show strong ventralizing activity and at high concentrations, it showed activation even in the absence of Dexamethasone. Injection of concentrations with no significant background level activity did not affect NC migration after treatment with Dexamethasone at premigratory or migratory NC cells stages (data not shown). D Graph summarizing the percentage of ventralized embryos of 3 independent experiments in embryos injected with increasing concentrations of Tcf3∆C-GR RNA. Numbers of injected embryos and standard error of the means are indicated for each column. Tcf3∆C-GR is a potent ventralizing agent, however, also active to some extent in the absence of Dexamethasone. Therefore, we used 50 pg for our migration assays, as background effects are comparably low at this concentration Image published in: Maj E et al. (2016) Copyright © 2016. Image reproduced with permission of the Publisher, Elsevier B. V. Larger Image Printer Friendly View

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