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. 2017 Dec 7;8:1973. doi: 10.1038/s41467-017-02076-0

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© The Author(s) 2017

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 Commonslicense, unless indicated otherwise in a credit line to the material. If material is not included in the article’sCreative 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. 9 — Frzb and the Wnt8–Frzb complex accumulates on N-acetyl-rich HS clusters. Intercellular distributions of HS and Wnt8 or Frzb in the Xenopus embryo (st. 10.5) were visualized by immunostaining (1st to 3rd panels from the left). Normalized intensities of Frzb-HAi or Wnt8-Myc (green) and HS (magenta) along the cell boundaries were plotted in graphs (4th panels). The start and end points are indicated by yellow and cyan arrowheads, respectively, in the merged pictures. Paired sets of normalized intensities of N-acetyl and N-sulfo signals at each position were plotted (5th panels). Values of calculated correlation coefficients (r) are shown in the graphs. a Frzb-HAi. Internally HA-tagged Frzb (Frzb-HAi) preferentially colocalized with N-acetyl HS compared to N-sulfo HS. b Wnt8-Myc. Conversely, Wnt8-Myc preferentially colocalized with N-sulfo HS compared to N-acetyl HS. c Wnt8-Myc in the presence of Frzb. When Frzb was expressed adjacently, Wnt8-Myc became colocalized preferentially to N-acetyl HS similar to Frzb-HAi. Images are a representative of four independent experiments. Amounts of mRNAs (ng/embryo): frzb-HAi 0.25; wnt8-myc, 0.25; for frzb, 1.0. Scale bars, 20 μm. a.u., arbitrary units