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. 2020 Jan 21;11:414. doi: 10.1038/s41467-019-14149-3

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© The Author(s) 2020

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 3 — a The model of the active-like FZD₆ (blue) showing a pronounced outward-motion (Δ) of the TM6 as compared to the inactive model (gray), justifying positioning of FRET acceptor and donor in the ICL3 and C-terminus, respectively. b The scheme depicts the FZD₆–FlAsH–TFP construct. The FlAsH-binding motif (FLNCCPGCCMEP) was inserted into the ICL3, between G404 and R405. Receptor activation is predicted to result in a loss of FRET due to conformational rearrangement in accordance to previous data obtained for FZD₅¹⁶. c WNT-5A induced a concentration-dependent decrease of the FRET ratio (FlAsH/TFP) in HEK293 cells overexpressing FZD₆–FlAsH–TFP. The FRET ratio change induced by each concentration was normalized to basal FRET ratio. Data are represented as mean ± s.e.m. of total n = 5 individual experiments. d SAG1.3 induces a concentration-dependent decrease of the FRET ratio (FlAsH/TFP) in HEK293 cells overexpressing FZD₆–FlAsH–TFP. The FRET ratio change induced by each concentration was normalized to basal FRET ratio. Data are presented as mean ± s.e.m. of total n = 7 individual experiments. Source data are provided as a Source Data file.