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. 2015 Nov 24;10:5. doi: 10.5334/1750-2187-10-6

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Copyright: © 2015 The Author(s)

This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 Unported License (CC-BY 3.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. See http://creativecommons.org/licenses/by/3.0/.

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Characterization of Frizzled-4 variants implicated in FEVR. (A) A snake diagram of the V5-tagged mouse Frizzled-4 (V5mFz4) used in this study. The V5 epitope cloned into the amino terminus after the signal peptide is represented by a red shading. The putative CRD domain or ligand binding site is colored blue. The general localization of amino acids found to be mutated in certain FEVR patients that were investigated in this study are shown inside stars. F328 of iloop2 is displayed in an orange star. (B) V5mFz4 variants were overexpressed in HEK293 cells with hLRP5 and M50 reporter plasmid, stimulated with Norrin and Lef/Tcf-dependent luciferase activity was assayed as described. (C) IIF of nonpermeabilized HeLa cells transfected with V5mFz4 variants using a V5 primary antibody.