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. 2019 Apr 30;9:6726. doi: 10.1038/s41598-019-43143-4

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© The Author(s) 2019

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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ANXA7 is required for ALG-2 binding to supported membranes. (a) Schematic representation of supported membrane model composed of primary and secondary membranes. Non-vesicular membrane patches with open edges were used in the subsequent experiments (POPC/POPS, 9:1 molar ratio). (b) Representative sequential images before and after addition of recombinant ANXA7-GFP protein to a membrane patch stained with DiD (DiD channel: upper panel. Lower panel: GFP channel) in the presence of Ca²⁺ (Also see Supplementary Movie S2). Note, ANXA7-GFP induces a pearl-like membrane conformation. (c) Similar experiment as in b but in the absence of Ca²⁺ (top panel) or POPS (lower panel). (d) Sequential images before and after addition of recombinant ALG-2-GFP protein to a membrane patch stained with DiD in the presence of Ca²⁺. (e) Addition of ANXA7 (without GFP) and ALG-2-GFP proteins trigger ALG-2-GFP membrane binding. Accumulation of ALG-2-GFP into pearl-like peripheral structures induced by ANXA7 are indicated by yellow arrows (upper panel, DiD. Lower panel: GFP channel).