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The assay reconstitutes actomyosin motion via myosin ID interacting dynamically with lipid membranes, enabling quantitative measurement of actin gliding velocity on the inner surface of phospholipid membranes.",{"@graph":14,"@context":72},[15,34,55],{"@type":16,"itemListElement":17},"BreadcrumbList",[18,23,27,31],{"item":19,"name":20,"@type":21,"position":22},"https://docshare.wps.com","Home","ListItem",1,{"item":24,"name":25,"@type":21,"position":26},"https://docshare.wps.com/document/","Document",2,{"item":28,"name":29,"@type":21,"position":30},"https://docshare.wps.com/document/research-report/","Research & 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approach?",{"text":71,"@type":63},"W/O emulsion droplets provide confined geometry that mimics the intracellular environment and supports dynamic interaction between actomyosin and lipid membranes.","https://schema.org",{"og:url":32,"og:type":74,"og:title":10,"og:site_name":45,"og:description":12},"article",{"robots":76,"canonical":32},"index,follow",{"doc_id":78,"site_id":7},443944,1790768093,{"code":4,"msg":81,"data":82},"success",[83,87,91,95,100,105,110,114,119,122,126],{"id":22,"doc_module":4,"doc_module_name":25,"category_name":84,"show_sort_weight":85,"slug":86},"Story & 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www.elsevier.com/locate/methodsx)[ www.elsevier.com/locate/methodsx](homepage: www.elsevier.com/locate/methodsx)  \n| A method for reconstituting the motility of membrane-bound myosin on the surface of the cell-sized W/O droplet | |\n| --- | --- |\n| Yusei Satoa,b,*, Rieko Sumiyoshi a, Masahito Hayashi c, Masahiko Yamagishi a,d, Junichiro Yajima a,d,e,* \u003Cbr>a Department of Life Sciences, Graduate School of Arts and Sciences, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan b Institute for Extra-cutting-edge Science and Technology Avant-garde Research (X-star), Japan Agency for Marine-Earth Science and Technology (JAMSTEC), 2-15 Natsushima-cho, Yokosuka, Kanagawa 237-0061, Japan\u003Cbr>c Department of Biotechnology and Life Science, Tokyo University of Agriculture and Technology, 2-24-16 Naka-cho, Koganei, Tokyo 184-8588, Japan\u003Cbr>d Komaba Institute for Science, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan\u003Cbr>e Research Center for Complex Systems Biology, Universal Biology Institute, The University of Tokyo, 3-8-1 Komaba, Meguro-ku, Tokyo 153-8902, Japan |  |\n| G R A P H I C A L A B S T R A C T |  |\n| |  |\n\nA R T I C L E I N F O  \nKeywords: Myosin  \nA B S T R A C T  \nMembrane-bound myosin generates force through interactions with the cytoskeletal actin filament beneath the cell membrane and constitutes the mechanical basis for living cells. Myosin ID,  \nFor a published article:Yusei Sato, Rieko Sumiyoshi, Masahiko Yamagishi, Takeshi Haraguchi, Kyohei Matsuda, Suguru Sato, Kohji Ito, Junichiro YajimaReconstructing the Motility Driven by Membrane-Bound Myosin on the Inner Surface of Cell-Sized Droplets Langmuir, 41 (2025), pp. 10,077–10,084 [https://doi.org/10.1021/acs.langmuir.4c04123](https://doi.org/10.1021/acs.langmuir.4c04123)  \n* Corresponding authors.  \nE-mail addresses: [yuseisato@jamstec.go.jp](yuseisato@jamstec.go.jp) (Y. Sato), [yajima@bio.c.u-tokyo.ac.jp](yajima@bio.c.u-tokyo.ac.jp) (J. Yajima).  \n[https://doi.org/10.1016/j.mex.2025.103755](https://doi.org/10.1016/j.mex.2025.103755)  \nReceived 15 October 2025; Accepted 9 December 2025  \nAvailable online 9 December 2025  \n2215-0161/© 2025 The Authors. Published by Elsevier B.V. This is an open access article under the CC BY license  \n([http://creativecommons.org/licenses/by/4.0/](http://creativecommons.org/licenses/by/4.0/)).  \nY. Sato et al. MethodsX 16 (2026) 103755  \nActin Artificial cell Motility  \nCell membrane Phospholipid  \na membrane-bound myosin, drives the gliding motion of actin filaments and binds to phospholipids in the lipid membrane of a living cell. Here, we describe the in-droplet actin filament gliding assay, a method that reconstitutes the motility of actomyosin that dynamically interacts with lipid membranes within water-in-oil (W/O) droplets, which mimic the confined geometry of the intracellular environment. Our method enables quantification of the gliding velocity of actin filaments driven by myosin ID on the inner surface ofW/O droplets surrounded by a phospholipid membrane. The in-droplet actin filament gliding assay provides a valuable platform for reconstituting the motile properties of other membrane-bound myosins on membrane surfaces in confined spaces and for analyzing the dynamics of actomyosin networks. The main features and applications of this method are as follows:  \n• Reconstitution of actin filament gliding driven by membrane-bound myosin ID within confined water-in-oil droplets.  \n• Quantitative evaluation of actomyosin dynamics on the inner surface of the lipid membrane within water-in-oil-droplets.  \n• Broadly applicable assay platform for studying the motile properties of membrane-associated myosin families.  \n\n| Specifications table |  |\n| --- | --- |\n| Subject area | Biochemistry, Genetics and Molecular Biology |\n| More specific subject area | Motility assay for motor prote","cbCaiaLVBexFNGeA","https://ap.wps.com/l/cbCaiaLVBexFNGeA","pdf",3228003,"English","# Background\n## Myosin–actin motility and gliding assays\n## Geometry-mimicking platforms: supported lipid bilayers and W/O droplets\n# Method overview\n## In-droplet actin filament gliding assay concept\n## Quantitative readout of gliding velocity\n# Main features and applications","[{\"question\":\"What method does the article introduce for studying membrane-bound myosin motility?\",\"answer\":\"It describes an in-droplet actin filament gliding assay performed inside water-in-oil (W/O) droplets with a surrounding phospholipid membrane.\"},{\"question\":\"How does the assay quantify actomyosin behavior?\",\"answer\":\"It enables quantitative evaluation of actin filament gliding velocity driven by myosin ID on the inner surface of the lipid membrane within W/O droplets.\"},{\"question\":\"Why are water-in-oil droplets used in this reconstitution approach?\",\"answer\":\"W/O emulsion droplets provide confined geometry that mimics the intracellular environment and supports dynamic interaction between actomyosin and lipid membranes.\"}]","A method for reconstituting the motility of membrane-bound myosin on the surface of the cell-sized W/O droplet | PDF",1790706172,23]