{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,1]],"date-time":"2026-07-01T18:55:51Z","timestamp":1782932151737,"version":"3.54.5"},"reference-count":84,"publisher":"Springer Science and Business Media LLC","issue":"1","license":[{"start":{"date-parts":[[2022,12,7]],"date-time":"2022-12-07T00:00:00Z","timestamp":1670371200000},"content-version":"tdm","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"},{"start":{"date-parts":[[2022,12,7]],"date-time":"2022-12-07T00:00:00Z","timestamp":1670371200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0"}],"content-domain":{"domain":["link.springer.com"],"crossmark-restriction":false},"short-container-title":["Commun Biol"],"abstract":"<jats:title>Abstract<\/jats:title><jats:p>Physical interactions of cells with the underlying extracellular matrix (ECM) play key roles in multiple cellular processes. The actin cytoskeleton is a central driver and regulator of cellular dynamics, that produces membrane-protrusions such as lamellipodia and filopodia. Here, we examined actin organization in expanding lamellipodia during early stages of cell spreading. To gain insight into the 3D actin organization, we plated fibroblasts on galectin-8 coated EM grids, an ECM protein presents in disease states. We then combined cryo-electron tomography with advanced image processing tools for reconstructing the structure of F-actin in the lamellipodia. This approach enabled us to resolve the polarity and orientation of filaments, and the structure of the Arp2\/3 complexes associated with F-actin branches. We show that F-actin in lamellipodial protrusions forms a dense network with three distinct sub-domains. One consists primarily of radial filaments, with their barbed ends pointing towards the membrane, the other is enriched with parallel filaments that run between the radial fibers, in addition to an intermediate sub-domain. Surprisingly, a minor, yet significant (~10%) population of actin filaments, are oriented with their barbed-ends towards the cell center. Our results provide structural insights into F-actin assembly and dynamic reorganization in the leading\u00a0edge of spreading cells.<\/jats:p>","DOI":"10.1038\/s42003-022-04288-7","type":"journal-article","created":{"date-parts":[[2022,12,7]],"date-time":"2022-12-07T02:41:56Z","timestamp":1670380916000},"update-policy":"https:\/\/doi.org\/10.1007\/springer_crossmark_policy","source":"Crossref","is-referenced-by-count":15,"title":["A network of mixed actin polarity in the leading edge of spreading cells"],"prefix":"10.1038","volume":"5","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-7411-9210","authenticated-orcid":false,"given":"Wen-Lu","family":"Chung","sequence":"first","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0638-9347","authenticated-orcid":false,"given":"Matthias","family":"Eibauer","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Wenhong","family":"Li","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Rajaa","family":"Boujemaa-Paterski","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8559-6373","authenticated-orcid":false,"given":"Benjamin","family":"Geiger","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-0994-2937","authenticated-orcid":false,"given":"Ohad","family":"Medalia","sequence":"additional","affiliation":[],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"297","published-online":{"date-parts":[[2022,12,7]]},"reference":[{"key":"4288_CR1","doi-asserted-by":"publisher","first-page":"1973","DOI":"10.1073\/pnas.1119578109","volume":"109","author":"K Tanner","year":"2012","unstructured":"Tanner, K., Mori, H., Mroue, R., Bruni-Cardoso, A. & Bissell, M. J. Coherent angular motion in the establishment of multicellular architecture of glandular tissues. Proc. Natl Acad. Sci. USA 109, 1973\u20131978 (2012).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"4288_CR2","doi-asserted-by":"publisher","first-page":"a023234","DOI":"10.1101\/cshperspect.a023234","volume":"9","author":"AI Bachir","year":"2017","unstructured":"Bachir, A. I., Horwitz, A. R., Nelson, W. J. & Bianchini, J. M. Actin-based adhesion modules mediate cell interactions with the extracellular matrix and neighboring cells. Cold Spring Harb. Perspect. Biol. 9, a023234 (2017).","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"4288_CR3","doi-asserted-by":"publisher","first-page":"1","DOI":"10.1016\/bs.ctdb.2018.02.002","volume":"130","author":"JM Muncie","year":"2018","unstructured":"Muncie, J. M. & Weaver, V. M. The physical and biochemical properties of the extracellular matrix regulate cell fate. Curr. Top. Dev. Biol. 130, 1\u201337 (2018).","journal-title":"Curr. Top. Dev. Biol."},{"key":"4288_CR4","doi-asserted-by":"publisher","first-page":"a018267","DOI":"10.1101\/cshperspect.a018267","volume":"10","author":"T Svitkina","year":"2018","unstructured":"Svitkina, T. The actin cytoskeleton and actin-based motility. Cold Spring Harb. Perspect. Biol. 10, a018267 (2018).","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"4288_CR5","doi-asserted-by":"publisher","first-page":"633","DOI":"10.1038\/nrm2957","volume":"11","author":"JT Parsons","year":"2010","unstructured":"Parsons, J. T., Horwitz, A. R. & Schwartz, M. A. Cell adhesion: integrating cytoskeletal dynamics and cellular tension. Nat. Rev. Mol. Cell Biol. 11, 633\u2013643 (2010).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"4288_CR6","doi-asserted-by":"publisher","first-page":"694","DOI":"10.1016\/j.cub.2007.02.058","volume":"17","author":"D Cuvelier","year":"2007","unstructured":"Cuvelier, D. et al. The universal dynamics of cell spreading. Curr. Biol. 17, 694\u2013699 (2007).","journal-title":"Curr. Biol."},{"key":"4288_CR7","doi-asserted-by":"publisher","first-page":"453","DOI":"10.1016\/S0092-8674(03)00120-X","volume":"112","author":"TD Pollard","year":"2003","unstructured":"Pollard, T. D. & Borisy, G. G. Cellular motility driven by assembly and disassembly of actin filaments. Cell 112, 453\u2013465 (2003).","journal-title":"Cell"},{"key":"4288_CR8","doi-asserted-by":"publisher","first-page":"69","DOI":"10.1016\/j.ceb.2020.05.008","volume":"66","author":"M Kelkar","year":"2020","unstructured":"Kelkar, M., Bohec, P. & Charras, G. Mechanics of the cellular actin cortex: from signalling to shape change. Curr. Opin. Cell Biol. 66, 69\u201378 (2020).","journal-title":"Curr. Opin. Cell Biol."},{"key":"4288_CR9","doi-asserted-by":"publisher","first-page":"R1063","DOI":"10.1016\/j.cub.2010.10.059","volume":"20","author":"A Byron","year":"2010","unstructured":"Byron, A., Morgan, M. R. & Humphries, M. J. Adhesion signalling complexes. Curr. Biol. 20, R1063\u2013R1067 (2010).","journal-title":"Curr. Biol."},{"key":"4288_CR10","doi-asserted-by":"publisher","first-page":"2508","DOI":"10.1016\/j.bpj.2014.10.041","volume":"107","author":"H Wolfenson","year":"2014","unstructured":"Wolfenson, H., Iskratsch, T. & Sheetz, M. P. Early events in cell spreading as a model for quantitative analysis of biomechanical events. Biophys. J. 107, 2508\u20132514 (2014).","journal-title":"Biophys. J."},{"key":"4288_CR11","doi-asserted-by":"publisher","first-page":"306","DOI":"10.1038\/ncb1692","volume":"10","author":"SA Koestler","year":"2008","unstructured":"Koestler, S. A., Auinger, S., Vinzenz, M., Rottner, K. & Small, J. V. Differentially oriented populations of actin filaments generated in lamellipodia collaborate in pushing and pausing at the cell front. Nat. Cell Biol. 10, 306\u2013313 (2008).","journal-title":"Nat. Cell Biol."},{"key":"4288_CR12","doi-asserted-by":"publisher","first-page":"483","DOI":"10.1042\/EBC20190015","volume":"63","author":"M Schaks","year":"2019","unstructured":"Schaks, M., Giannone, G. & Rottner, K. Actin dynamics in cell migration. Essays Biochem. 63, 483\u2013495 (2019).","journal-title":"Essays Biochem."},{"key":"4288_CR13","doi-asserted-by":"publisher","first-page":"33","DOI":"10.1016\/j.biochi.2016.09.019","volume":"130","author":"J Lemiere","year":"2016","unstructured":"Lemiere, J., Valentino, F., Campillo, C. & Sykes, C. How cellular membrane properties are affected by the actin cytoskeleton. Biochimie 130, 33\u201340 (2016).","journal-title":"Biochimie"},{"key":"4288_CR14","doi-asserted-by":"publisher","first-page":"472","DOI":"10.1016\/j.ceb.2006.08.012","volume":"18","author":"A Bershadsky","year":"2006","unstructured":"Bershadsky, A., Kozlov, M. & Geiger, B. Adhesion-mediated mechanosensitivity: a time to experiment, and a time to theorize. Curr. Opin. Cell Biol. 18, 472\u2013481 (2006).","journal-title":"Curr. Opin. Cell Biol."},{"key":"4288_CR15","doi-asserted-by":"publisher","first-page":"21","DOI":"10.1038\/nrm2593","volume":"10","author":"B Geiger","year":"2009","unstructured":"Geiger, B., Spatz, J. P. & Bershadsky, A. D. Environmental sensing through focal adhesions. Nat. Rev. Mol. Cell Biol. 10, 21\u201333 (2009).","journal-title":"Nat. Rev. Mol. Cell Biol."},{"key":"4288_CR16","doi-asserted-by":"publisher","first-page":"R667","DOI":"10.1016\/j.cub.2021.03.031","volume":"31","author":"DN Clarke","year":"2021","unstructured":"Clarke, D. N. & Martin, A. C. Actin-based force generation and cell adhesion in tissue morphogenesis. Curr. Biol. 31, R667\u2013R680 (2021).","journal-title":"Curr. Biol."},{"key":"4288_CR17","doi-asserted-by":"publisher","first-page":"1009","DOI":"10.1083\/jcb.145.5.1009","volume":"145","author":"TM Svitkina","year":"1999","unstructured":"Svitkina, T. M. & Borisy, G. G. Arp2\/3 complex and actin depolymerizing factor\/cofilin in dendritic organization and treadmilling of actin filament array in lamellipodia. J. Cell Biol. 145, 1009\u20131026 (1999).","journal-title":"J. Cell Biol."},{"key":"4288_CR18","doi-asserted-by":"publisher","first-page":"306","DOI":"10.1038\/35060104","volume":"3","author":"KJ Amann","year":"2001","unstructured":"Amann, K. J. & Pollard, T. D. The Arp2\/3 complex nucleates actin filament branches from the sides of pre-existing filaments. Nat. Cell Biol. 3, 306\u2013310 (2001).","journal-title":"Nat. Cell Biol."},{"key":"4288_CR19","doi-asserted-by":"publisher","first-page":"973","DOI":"10.1016\/j.cell.2011.12.034","volume":"148","author":"C Wu","year":"2012","unstructured":"Wu, C. et al. Arp2\/3 is critical for lamellipodia and response to extracellular matrix cues but is dispensable for chemotaxis. Cell 148, 973\u2013987 (2012).","journal-title":"Cell"},{"key":"4288_CR20","doi-asserted-by":"publisher","first-page":"112","DOI":"10.1016\/S0962-8924(01)02237-1","volume":"12","author":"JV Small","year":"2002","unstructured":"Small, J. V., Stradal, T., Vignal, E. & Rottner, K. The lamellipodium: where motility begins. Trends Cell Biol. 12, 112\u2013120 (2002).","journal-title":"Trends Cell Biol."},{"key":"4288_CR21","doi-asserted-by":"publisher","first-page":"1591","DOI":"10.1016\/S0006-3495(03)74969-8","volume":"84","author":"A Mogilner","year":"2003","unstructured":"Mogilner, A. & Oster, G. Force generation by actin polymerization II: the elastic ratchet and tethered filaments. Biophys. J. 84, 1591\u20131605 (2003).","journal-title":"Biophys. J."},{"key":"4288_CR22","doi-asserted-by":"publisher","first-page":"911","DOI":"10.1016\/S0091-679X(07)84029-5","volume":"84","author":"AE Carlsson","year":"2008","unstructured":"Carlsson, A. E. & Sept, D. Mathematical modeling of cell migration. Methods Cell Biol. 84, 911\u2013937 (2008).","journal-title":"Methods Cell Biol."},{"key":"4288_CR23","doi-asserted-by":"publisher","first-page":"1148","DOI":"10.1038\/s41556-021-00786-8","volume":"23","author":"A Mehidi","year":"2021","unstructured":"Mehidi, A. et al. Forces generated by lamellipodial actin filament elongation regulate the WAVE complex during cell migration. Nat. Cell Biol. 23, 1148\u20131162 (2021).","journal-title":"Nat. Cell Biol."},{"key":"4288_CR24","doi-asserted-by":"publisher","first-page":"397","DOI":"10.1083\/jcb.139.2.397","volume":"139","author":"TM Svitkina","year":"1997","unstructured":"Svitkina, T. M., Verkhovsky, A. B., McQuade, K. M. & Borisy, G. G. Analysis of the actin-myosin II system in fish epidermal keratocytes: mechanism of cell body translocation. J. Cell Biol. 139, 397\u2013415 (1997).","journal-title":"J. Cell Biol."},{"key":"4288_CR25","doi-asserted-by":"publisher","first-page":"4667","DOI":"10.1091\/mbc.e02-10-0630","volume":"14","author":"AB Verkhovsky","year":"2003","unstructured":"Verkhovsky, A. B. et al. Orientational order of the lamellipodial actin network as demonstrated in living motile cells. Mol. Biol. Cell 14, 4667\u20134675 (2003).","journal-title":"Mol. Biol. Cell"},{"key":"4288_CR26","first-page":"2775","volume":"125","author":"M Vinzenz","year":"2012","unstructured":"Vinzenz, M. et al. Actin branching in the initiation and maintenance of lamellipodia. J. Cell Sci. 125, 2775\u20132785 (2012).","journal-title":"J. Cell Sci."},{"key":"4288_CR27","doi-asserted-by":"publisher","first-page":"188","DOI":"10.1016\/j.cell.2017.07.051","volume":"171","author":"J Mueller","year":"2017","unstructured":"Mueller, J. et al. Load adaptation of lamellipodial actin networks. Cell 171, 188\u2013200.e116 (2017).","journal-title":"Cell"},{"key":"4288_CR28","doi-asserted-by":"publisher","first-page":"2731","DOI":"10.1002\/bit.24921","volume":"110","author":"AK Chen","year":"2013","unstructured":"Chen, A. K. et al. Cell spreading and proliferation in response to the composition and mechanics of engineered fibrillar extracellular matrices. Biotechnol. Bioeng. 110, 2731\u20132741 (2013).","journal-title":"Biotechnol. Bioeng."},{"key":"4288_CR29","doi-asserted-by":"publisher","first-page":"1397","DOI":"10.1038\/cr.2017.108","volume":"27","author":"D Wu","year":"2017","unstructured":"Wu, D. et al. Pairing of integrins with ECM proteins determines migrasome formation. Cell Res. 27, 1397\u20131400 (2017).","journal-title":"Cell Res."},{"key":"4288_CR30","doi-asserted-by":"publisher","first-page":"31956","DOI":"10.1074\/jbc.M401409200","volume":"279","author":"J Jokinen","year":"2004","unstructured":"Jokinen, J. et al. Integrin-mediated cell adhesion to type I collagen fibrils. J. Biol. Chem. 279, 31956\u201331963 (2004).","journal-title":"J. Biol. Chem."},{"key":"4288_CR31","doi-asserted-by":"publisher","first-page":"901","DOI":"10.1083\/jcb.200910095","volume":"189","author":"A Hotta","year":"2010","unstructured":"Hotta, A. et al. Laminin-based cell adhesion anchors microtubule plus ends to the epithelial cell basal cortex through LL5alpha\/beta. J. Cell Biol. 189, 901\u2013917 (2010).","journal-title":"J. Cell Biol."},{"key":"4288_CR32","doi-asserted-by":"publisher","first-page":"909","DOI":"10.1038\/ncb2095","volume":"12","author":"I Patla","year":"2010","unstructured":"Patla, I. et al. Dissecting the molecular architecture of integrin adhesion sites by cryo-electron tomography. Nat. Cell Biol. 12, 909\u2013915 (2010).","journal-title":"Nat. Cell Biol."},{"key":"4288_CR33","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-021-21142-2","volume":"12","author":"LS Fischer","year":"2021","unstructured":"Fischer, L. S. et al. Quantitative single-protein imaging reveals molecular complex formation of integrin, talin, and kindlin during cell adhesion. Nat. Commun. 12, 919 (2021).","journal-title":"Nat. Commun."},{"key":"4288_CR34","doi-asserted-by":"publisher","first-page":"e17","DOI":"10.1017\/S1462399408000719","volume":"10","author":"RY Yang","year":"2008","unstructured":"Yang, R. Y., Rabinovich, G. A. & Liu, F. T. Galectins: structure, function and therapeutic potential. Expert Rev. Mol. Med. 10, e17 (2008).","journal-title":"Expert Rev. Mol. Med."},{"key":"4288_CR35","doi-asserted-by":"publisher","DOI":"10.1038\/s41598-018-29843-3","volume":"8","author":"MS Tsai","year":"2018","unstructured":"Tsai, M. S. et al. Galectin-1 restricts vascular smooth muscle cell motility via modulating adhesion force and focal adhesion dynamics. Sci. Rep. 8, 11497 (2018).","journal-title":"Sci. Rep."},{"key":"4288_CR36","doi-asserted-by":"publisher","first-page":"2934","DOI":"10.3390\/ijms19102934","volume":"19","author":"A Bartolazzi","year":"2018","unstructured":"Bartolazzi, A. Galectins in cancer and translational medicine: from bench to bedside. Int. J. Mol. Sci. 19, 2934 (2018).","journal-title":"Int. J. Mol. Sci."},{"key":"4288_CR37","doi-asserted-by":"publisher","first-page":"907","DOI":"10.1093\/glycob\/cwu054","volume":"24","author":"MF Troncoso","year":"2014","unstructured":"Troncoso, M. F. et al. Galectin-8: a matricellular lectin with key roles in angiogenesis. Glycobiology 24, 907\u2013914 (2014).","journal-title":"Glycobiology"},{"key":"4288_CR38","doi-asserted-by":"publisher","first-page":"31285","DOI":"10.1074\/jbc.M100340200","volume":"276","author":"Y Levy","year":"2001","unstructured":"Levy, Y. et al. Galectin-8 functions as a matricellular modulator of cell adhesion. J. Biol. Chem. 276, 31285\u201331295 (2001).","journal-title":"J. Biol. Chem."},{"key":"4288_CR39","doi-asserted-by":"publisher","first-page":"4187","DOI":"10.1007\/s00018-018-2856-2","volume":"75","author":"D Weinmann","year":"2018","unstructured":"Weinmann, D. et al. Galectin-8 induces functional disease markers in human osteoarthritis and cooperates with galectins-1 and -3. Cell Mol. Life Sci. 75, 4187\u20134205 (2018).","journal-title":"Cell Mol. Life Sci."},{"key":"4288_CR40","doi-asserted-by":"publisher","DOI":"10.1186\/s40659-016-0091-6","volume":"49","author":"C Metz","year":"2016","unstructured":"Metz, C. et al. Galectin-8 promotes migration and proliferation and prevents apoptosis in U87 glioblastoma cells. Biol. Res. 49, 33 (2016).","journal-title":"Biol. Res."},{"key":"4288_CR41","doi-asserted-by":"publisher","first-page":"e44400","DOI":"10.1371\/journal.pone.0044400","volume":"7","author":"S Diskin","year":"2012","unstructured":"Diskin, S. et al. Galectin-8 promotes cytoskeletal rearrangement in trabecular meshwork cells through activation of Rho signaling. PLoS ONE 7, e44400 (2012).","journal-title":"PLoS ONE"},{"key":"4288_CR42","doi-asserted-by":"publisher","first-page":"135","DOI":"10.1042\/BST20200677","volume":"50","author":"Y Zick","year":"2022","unstructured":"Zick, Y. Galectin-8, cytokines, and the storm. Biochem Soc. Trans. 50, 135\u2013149 (2022).","journal-title":"Biochem Soc. Trans."},{"key":"4288_CR43","doi-asserted-by":"publisher","first-page":"jcs252221","DOI":"10.1242\/jcs.252221","volume":"134","author":"W Li","year":"2021","unstructured":"Li, W. et al. Differential cellular responses to adhesive interactions with galectin-8- and fibronectin-coated substrates. J. Cell Sci. 134, jcs252221 (2021).","journal-title":"J. Cell Sci."},{"key":"4288_CR44","doi-asserted-by":"publisher","first-page":"1209","DOI":"10.1126\/science.1076184","volume":"298","author":"O Medalia","year":"2002","unstructured":"Medalia, O. et al. Macromolecular architecture in eukaryotic cells visualized by cryoelectron tomography. Science 298, 1209\u20131213 (2002).","journal-title":"Science"},{"key":"4288_CR45","doi-asserted-by":"publisher","DOI":"10.1038\/ncomms2128","volume":"3","author":"NE Reticker-Flynn","year":"2012","unstructured":"Reticker-Flynn, N. E. et al. A combinatorial extracellular matrix platform identifies cell-extracellular matrix interactions that correlate with metastasis. Nat. Commun. 3, 1122 (2012).","journal-title":"Nat. Commun."},{"key":"4288_CR46","doi-asserted-by":"publisher","first-page":"1048","DOI":"10.1093\/glycob\/cww066","volume":"26","author":"M Friedel","year":"2016","unstructured":"Friedel, M., Andre, S., Goldschmidt, H., Gabius, H. J. & Schwartz-Albiez, R. Galectin-8 enhances adhesion of multiple myeloma cells to vascular endothelium and is an adverse prognostic factor. Glycobiology 26, 1048\u20131058 (2016).","journal-title":"Glycobiology"},{"key":"4288_CR47","first-page":"4099","volume":"126","author":"N Elad","year":"2013","unstructured":"Elad, N. et al. The role of integrin-linked kinase in the molecular architecture of focal adhesions. J. Cell Sci. 126, 4099\u20134107 (2013).","journal-title":"J. Cell Sci."},{"key":"4288_CR48","doi-asserted-by":"publisher","first-page":"983","DOI":"10.1038\/nmeth.4405","volume":"14","author":"M Chen","year":"2017","unstructured":"Chen, M. et al. Convolutional neural networks for automated annotation of cellular cryo-electron tomograms. Nat. Methods 14, 983\u2013985 (2017).","journal-title":"Nat. Methods"},{"key":"4288_CR49","doi-asserted-by":"publisher","first-page":"488","DOI":"10.1016\/j.str.2020.12.014","volume":"29","author":"B Martins","year":"2021","unstructured":"Martins, B. et al. Unveiling the polarity of actin filaments by cryo-electron tomography. Structure 29, 488\u2013498.e4 (2021).","journal-title":"Structure"},{"key":"4288_CR50","doi-asserted-by":"publisher","first-page":"374","DOI":"10.1038\/s41586-022-05241-8","volume":"611","author":"W Oosterheert","year":"2022","unstructured":"Oosterheert, W., Klink, B. U., Belyy, A., Pospich, S. & Raunser, S. Structural basis of actin filament assembly and aging. Nature 611, 374\u2013379 (2022).","journal-title":"Nature"},{"key":"4288_CR51","doi-asserted-by":"publisher","first-page":"1285","DOI":"10.1073\/pnas.1211164110","volume":"110","author":"BA Smith","year":"2013","unstructured":"Smith, B. A., Daugherty-Clarke, K., Goode, B. L. & Gelles, J. Pathway of actin filament branch formation by Arp2\/3 complex revealed by single-molecule imaging. Proc. Natl Acad. Sci. USA 110, 1285\u20131290 (2013).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"4288_CR52","doi-asserted-by":"publisher","first-page":"703","DOI":"10.1016\/S0092-8674(01)00605-5","volume":"107","author":"JA Cooper","year":"2001","unstructured":"Cooper, J. A., Wear, M. A. & Weaver, A. M. Arp2\/3 complex: advances on the inner workings of a molecular machine. Cell 107, 703\u2013705 (2001).","journal-title":"Cell"},{"key":"4288_CR53","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-020-20286-x","volume":"11","author":"F Fassler","year":"2020","unstructured":"Fassler, F., Dimchev, G., Hodirnau, V. V., Wan, W. & Schur, F. K. M. Cryo-electron tomography structure of Arp2\/3 complex in cells reveals new insights into the branch junction. Nat. Commun. 11, 6437 (2020).","journal-title":"Nat. Commun."},{"key":"4288_CR54","doi-asserted-by":"publisher","first-page":"1211","DOI":"10.1016\/j.str.2019.05.009","volume":"27","author":"M Jasnin","year":"2019","unstructured":"Jasnin, M. et al. The architecture of traveling actin waves revealed by cryo-electron tomography. Structure 27, 1211\u20131223.e1215 (2019).","journal-title":"Structure"},{"key":"4288_CR55","doi-asserted-by":"publisher","first-page":"1605","DOI":"10.1002\/jcc.20084","volume":"25","author":"EF Pettersen","year":"2004","unstructured":"Pettersen, E. F. et al. UCSF chimera\u2014a visualization system for exploratory research and analysis. J. Comput. Chem. 25, 1605\u20131612 (2004).","journal-title":"J. Comput. Chem."},{"key":"4288_CR56","doi-asserted-by":"publisher","first-page":"e2105004118","DOI":"10.1073\/pnas.2105004118","volume":"118","author":"S Sorrentino","year":"2021","unstructured":"Sorrentino, S. et al. Structural analysis of receptors and actin polarity in platelet protrusions. Proc. Natl Acad. Sci. USA 118, e2105004118 (2021).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"4288_CR57","doi-asserted-by":"publisher","first-page":"6304","DOI":"10.1073\/pnas.0913730107","volume":"107","author":"J Weichsel","year":"2010","unstructured":"Weichsel, J. & Schwarz, U. S. Two competing orientation patterns explain experimentally observed anomalies in growing actin networks. Proc. Natl Acad. Sci. USA 107, 6304\u20136309 (2010).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"4288_CR58","doi-asserted-by":"crossref","first-page":"2329","DOI":"10.1242\/jcs.184507","volume":"129","author":"SJ King","year":"2016","unstructured":"King, S. J. et al. Lamellipodia are crucial for haptotactic sensing and response. J. Cell Sci. 129, 2329\u20132342 (2016).","journal-title":"J. Cell Sci."},{"key":"4288_CR59","doi-asserted-by":"publisher","first-page":"443","DOI":"10.1083\/jcb.201406102","volume":"208","author":"HE Johnson","year":"2015","unstructured":"Johnson, H. E. et al. F-actin bundles direct the initiation and orientation of lamellipodia through adhesion-based signaling. J. Cell Biol. 208, 443\u2013455 (2015).","journal-title":"J. Cell Biol."},{"key":"4288_CR60","doi-asserted-by":"publisher","first-page":"3589","DOI":"10.1529\/biophysj.105.065383","volume":"89","author":"E Atilgan","year":"2005","unstructured":"Atilgan, E., Wirtz, D. & Sun, S. X. Morphology of the lamellipodium and organization of actin filaments at the leading edge of crawling cells. Biophys. J. 89, 3589\u20133602 (2005).","journal-title":"Biophys. J."},{"key":"4288_CR61","doi-asserted-by":"publisher","first-page":"7086","DOI":"10.1073\/pnas.0701943104","volume":"104","author":"TE Schaus","year":"2007","unstructured":"Schaus, T. E., Taylor, E. W. & Borisy, G. G. Self-organization of actin filament orientation in the dendritic-nucleation\/array-treadmilling model. Proc. Natl Acad. Sci. USA 104, 7086\u20137091 (2007).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"4288_CR62","doi-asserted-by":"publisher","first-page":"e74389","DOI":"10.7554\/eLife.74389","volume":"11","author":"RM Garner","year":"2022","unstructured":"Garner, R. M. & Theriot, J. A. Leading edge maintenance in migrating cells is an emergent property of branched actin network growth. Elife 11, e74389 (2022).","journal-title":"Elife"},{"key":"4288_CR63","doi-asserted-by":"publisher","first-page":"e3234","DOI":"10.1371\/journal.pone.0003234","volume":"3","author":"AY Alexandrova","year":"2008","unstructured":"Alexandrova, A. Y. et al. Comparative dynamics of retrograde actin flow and focal adhesions: formation of nascent adhesions triggers transition from fast to slow flow. PLoS ONE 3, e3234 (2008).","journal-title":"PLoS ONE"},{"key":"4288_CR64","doi-asserted-by":"publisher","first-page":"3923","DOI":"10.1242\/jcs.095653","volume":"124","author":"M Vicente-Manzanares","year":"2011","unstructured":"Vicente-Manzanares, M. & Horwitz, A. R. Adhesion dynamics at a glance. J. Cell Sci. 124, 3923\u20133927 (2011).","journal-title":"J. Cell Sci."},{"key":"4288_CR65","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1093\/jb\/mvu053","volume":"156","author":"S Yamashiro","year":"2014","unstructured":"Yamashiro, S. & Watanabe, N. A new link between the retrograde actin flow and focal adhesions. J. Biochem. 156, 239\u2013248 (2014).","journal-title":"J. Biochem."},{"key":"4288_CR66","doi-asserted-by":"publisher","first-page":"9660","DOI":"10.1073\/pnas.0300552101","volume":"101","author":"P Vallotton","year":"2004","unstructured":"Vallotton, P., Gupton, S. L., Waterman-Storer, C. M. & Danuser, G. Simultaneous mapping of filamentous actin flow and turnover in migrating cells by quantitative fluorescent speckle microscopy. Proc. Natl Acad. Sci. USA 101, 9660\u20139665 (2004).","journal-title":"Proc. Natl Acad. Sci. USA"},{"key":"4288_CR67","doi-asserted-by":"publisher","first-page":"e53990","DOI":"10.7554\/eLife.53990","volume":"9","author":"R Boujemaa-Paterski","year":"2020","unstructured":"Boujemaa-Paterski, R. et al. Talin-activated vinculin interacts with branched actin networks to initiate bundles. Elife 9, e53990 (2020).","journal-title":"Elife"},{"key":"4288_CR68","doi-asserted-by":"publisher","first-page":"163","DOI":"10.1083\/jcb.201303129","volume":"202","author":"I Thievessen","year":"2013","unstructured":"Thievessen, I. et al. Vinculin-actin interaction couples actin retrograde flow to focal adhesions, but is dispensable for focal adhesion growth. J. Cell Biol. 202, 163\u2013177 (2013).","journal-title":"J. Cell Biol."},{"key":"4288_CR69","doi-asserted-by":"publisher","first-page":"1012","DOI":"10.1038\/ncb2321","volume":"13","author":"C Yang","year":"2011","unstructured":"Yang, C. & Svitkina, T. Visualizing branched actin filaments in lamellipodia by electron tomography. Nat. Cell Biol. 13, 1012\u20131013 (2011).","journal-title":"Nat. Cell Biol."},{"key":"4288_CR70","doi-asserted-by":"publisher","first-page":"239","DOI":"10.1083\/jcb.201112113","volume":"197","author":"P Suraneni","year":"2012","unstructured":"Suraneni, P. et al. The Arp2\/3 complex is required for lamellipodia extension and directional fibroblast cell migration. J. Cell Biol. 197, 239\u2013251 (2012).","journal-title":"J. Cell Biol."},{"key":"4288_CR71","doi-asserted-by":"publisher","DOI":"10.1038\/s41467-017-00455-1","volume":"8","author":"R Boujemaa-Paterski","year":"2017","unstructured":"Boujemaa-Paterski, R. et al. Network heterogeneity regulates steering in actin-based motility. Nat. Commun. 8, 655 (2017).","journal-title":"Nat. Commun."},{"key":"4288_CR72","doi-asserted-by":"publisher","first-page":"1039","DOI":"10.1038\/ncb1763","volume":"10","author":"CK Choi","year":"2008","unstructured":"Choi, C. K. et al. Actin and alpha-actinin orchestrate the assembly and maturation of nascent adhesions in a myosin II motor-independent manner. Nat. Cell Biol. 10, 1039\u20131050 (2008).","journal-title":"Nat. Cell Biol."},{"key":"4288_CR73","doi-asserted-by":"publisher","first-page":"1090","DOI":"10.1038\/nmeth.4431","volume":"14","author":"P Ringer","year":"2017","unstructured":"Ringer, P. et al. Multiplexing molecular tension sensors reveals piconewton force gradient across talin-1. Nat. Methods 14, 1090\u20131096 (2017).","journal-title":"Nat. Methods"},{"key":"4288_CR74","doi-asserted-by":"publisher","first-page":"263","DOI":"10.1038\/nature09198","volume":"466","author":"C Grashoff","year":"2010","unstructured":"Grashoff, C. et al. Measuring mechanical tension across vinculin reveals regulation of focal adhesion dynamics. Nature 466, 263\u2013266 (2010).","journal-title":"Nature"},{"key":"4288_CR75","doi-asserted-by":"publisher","first-page":"671","DOI":"10.1038\/nmeth.2089","volume":"9","author":"CA Schneider","year":"2012","unstructured":"Schneider, C. A., Rasband, W. S. & Eliceiri, K. W. NIH Image to ImageJ: 25 years of image analysis. Nat. Methods 9, 671\u2013675 (2012).","journal-title":"Nat. Methods"},{"key":"4288_CR76","doi-asserted-by":"publisher","first-page":"36","DOI":"10.1016\/j.jsb.2005.07.007","volume":"152","author":"DN Mastronarde","year":"2005","unstructured":"Mastronarde, D. N. Automated electron microscope tomography using robust prediction of specimen movements. J. Struct. Biol. 152, 36\u201351 (2005).","journal-title":"J. Struct. Biol."},{"key":"4288_CR77","doi-asserted-by":"publisher","first-page":"227","DOI":"10.1016\/j.jsb.2004.10.006","volume":"149","author":"S Nickell","year":"2005","unstructured":"Nickell, S. et al. TOM software toolbox: acquisition and analysis for electron tomography. J. Struct. Biol. 149, 227\u2013234 (2005).","journal-title":"J. Struct. Biol."},{"key":"4288_CR78","doi-asserted-by":"publisher","first-page":"488","DOI":"10.1016\/j.jsb.2012.09.008","volume":"180","author":"M Eibauer","year":"2012","unstructured":"Eibauer, M. et al. Unraveling the structure of membrane proteins in situ by transfer function corrected cryo-electron tomography. J. Struct. Biol. 180, 488\u2013496 (2012).","journal-title":"J. Struct. Biol."},{"key":"4288_CR79","doi-asserted-by":"publisher","first-page":"519","DOI":"10.1016\/j.jsb.2012.09.006","volume":"180","author":"SH Scheres","year":"2012","unstructured":"Scheres, S. H. RELION: implementation of a Bayesian approach to cryo-EM structure determination. J. Struct. Biol. 180, 519\u2013530 (2012).","journal-title":"J. Struct. Biol."},{"key":"4288_CR80","unstructured":"Arthur, D. & Vassilvitskii, S. k-means++: the advantages of careful seeding. In Proc. eighteenth annual ACM-SIAM symposium on Discrete algorithms 1027\u20131035 (Society for Industrial and Applied Mathematics, New Orleans, 2007)."},{"key":"4288_CR81","unstructured":"Poulsen, A. T. MATLAB Central File Exchange. https:\/\/www.mathworks.com\/matlabcentral\/fileexchange\/47950-mseb-x-y-errbar-lineprops-transparent. (2022)."},{"key":"4288_CR82","unstructured":"Campbell, R. notBoxPlot. (GitHub; 2022) https:\/\/www.mathworks.com\/matlabcentral\/fileexchange\/26508-notboxplot."},{"key":"4288_CR83","doi-asserted-by":"publisher","first-page":"177","DOI":"10.1016\/j.jsb.2011.12.003","volume":"178","author":"T Hrabe","year":"2012","unstructured":"Hrabe, T. et al. PyTom: a python-based toolbox for localization of macromolecules in cryo-electron tomograms and subtomogram analysis. J. Struct. Biol. 178, 177\u2013188 (2012).","journal-title":"J. Struct. Biol."},{"key":"4288_CR84","doi-asserted-by":"publisher","first-page":"71","DOI":"10.1006\/jsbi.1996.0013","volume":"116","author":"JR Kremer","year":"1996","unstructured":"Kremer, J. R., Mastronarde, D. N. & McIntosh, J. R. Computer visualization of three-dimensional image data using IMOD. J. Struct. Biol. 116, 71\u201376 (1996).","journal-title":"J. Struct. Biol."}],"container-title":["Communications Biology"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.nature.com\/articles\/s42003-022-04288-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s42003-022-04288-7","content-type":"text\/html","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/www.nature.com\/articles\/s42003-022-04288-7.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2024,10,10]],"date-time":"2024-10-10T00:32:06Z","timestamp":1728520326000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.nature.com\/articles\/s42003-022-04288-7"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,12,7]]},"references-count":84,"journal-issue":{"issue":"1","published-online":{"date-parts":[[2022,12]]}},"alternative-id":["4288"],"URL":"https:\/\/doi.org\/10.1038\/s42003-022-04288-7","relation":{},"ISSN":["2399-3642"],"issn-type":[{"value":"2399-3642","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,12,7]]},"assertion":[{"value":"1 November 2022","order":1,"name":"received","label":"Received","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"22 November 2022","order":2,"name":"accepted","label":"Accepted","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"7 December 2022","order":3,"name":"first_online","label":"First Online","group":{"name":"ArticleHistory","label":"Article History"}},{"value":"The authors declare no competing interests.","order":1,"name":"Ethics","group":{"name":"EthicsHeading","label":"Competing interests"}}],"article-number":"1338"}}