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Figure S6. In vitro analysis of Wnt8 binding to Noggin4 by quantitative co-immunoprecipitation and Surface Plasmon Resonance assay. (a) Representative examples of Western blotting (WB) with antiFlag-antibody of indicated amounts of Flag-BAP (loading standard), Flag-Wnt8 eluted by Myc-peptide after incubation with Myc-Resin+Myc-Noggin4, Flag-Wnt8 eluted by Myc-peptide after incubation with Myc-resin+control samples and unbound Flag-Wnt8 after incubation with Myc-Resin+Myc-Noggin4. Loading standards and experimental samples were put on the same gels. (b) Adsorption isotherms drawn over experimental points (Table S2) obtained from WB images. Theoretical fits based on equations (3) and (4) are depicted with solid and dashed lines, respectively. Parameter values are provided in Materials and Methods. (c) Titration curves obtained from SPR measurements for interaction between immobilised Noggin4 and Wnt8. Image published in: Eroshkin FM et al. (2016) Copyright © 2016, Macmillan Publishers Limited. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution license Larger Image Printer Friendly View

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