{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,3,29]],"date-time":"2026-03-29T16:46:45Z","timestamp":1774802805037,"version":"3.50.1"},"reference-count":40,"publisher":"Public Library of Science (PLoS)","issue":"5","license":[{"start":{"date-parts":[[2021,5,11]],"date-time":"2021-05-11T00:00:00Z","timestamp":1620691200000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"DOI":"10.13039\/501100005363","name":"Universidad de Buenos Aires","doi-asserted-by":"publisher","award":["20020170100482BA"],"award-info":[{"award-number":["20020170100482BA"]}],"id":[{"id":"10.13039\/501100005363","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005363","name":"Universidad de Buenos Aires","doi-asserted-by":"publisher","award":["20020130300054BA"],"award-info":[{"award-number":["20020130300054BA"]}],"id":[{"id":"10.13039\/501100005363","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100005363","name":"Universidad de Buenos Aires","doi-asserted-by":"publisher","award":["20020120100155BA"],"award-info":[{"award-number":["20020120100155BA"]}],"id":[{"id":"10.13039\/501100005363","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003074","name":"Agencia Nacional de Promoci\u00f3n Cient\u00edfica y Tecnol\u00f3gica","doi-asserted-by":"publisher","award":["PICT 2010-0457"],"award-info":[{"award-number":["PICT 2010-0457"]}],"id":[{"id":"10.13039\/501100003074","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["www.plosone.org"],"crossmark-restriction":false},"short-container-title":["PLoS ONE"],"abstract":"<jats:p>Cells exert traction forces on the extracellular matrix to which they are adhered through the formation of focal adhesions. Spatial-temporal regulation of traction forces is crucial in cell adhesion, migration, cellular division, and remodeling of the extracellular matrix. By cultivating cells on polyacrylamide hydrogels of different stiffness we were able to investigate the effects of substrate stiffness on the generation of cellular traction forces by Traction Force Microscopy (TFM), and characterize the molecular dynamics of the focal adhesion protein zyxin by Fluorescence Correlation Spectroscopy (FCS) and Fluorescence Recovery After Photobleaching (FRAP). As the rigidity of the substrate increases, we observed an increment of both, cellular traction generation and zyxin residence time at the focal adhesions, while its diffusion would not be altered. Moreover, we found a positive correlation between the traction forces exerted by cells and the residence time of zyxin at the substrate elasticities studied. We found that this correlation persists at the subcellular level, even if there is no variation in substrate stiffness, revealing that focal adhesions that exert greater traction present longer residence time for zyxin, i.e., zyxin protein has less probability to dissociate from the focal adhesion.<\/jats:p>","DOI":"10.1371\/journal.pone.0251411","type":"journal-article","created":{"date-parts":[[2021,5,11]],"date-time":"2021-05-11T17:27:13Z","timestamp":1620754033000},"page":"e0251411","update-policy":"https:\/\/doi.org\/10.1371\/journal.pone.corrections_policy","source":"Crossref","is-referenced-by-count":8,"title":["Correlation of cellular traction forces and dissociation kinetics of adhesive protein zyxin revealed by multi-parametric live cell microscopy"],"prefix":"10.1371","volume":"16","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-9417-9600","authenticated-orcid":true,"given":"Lorena","family":"Sigaut","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-7887-075X","authenticated-orcid":true,"given":"Micaela","family":"Bianchi","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Catalina","family":"von Bilderling","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2714-3162","authenticated-orcid":true,"given":"L\u00eda Isabel","family":"Pietrasanta","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"340","published-online":{"date-parts":[[2021,5,11]]},"reference":[{"issue":"1","key":"pone.0251411.ref001","doi-asserted-by":"crossref","first-page":"21","DOI":"10.1038\/nrm2593","article-title":"Environmental sensing through focal adhesions","volume":"10","author":"B Geiger","year":"2009","journal-title":"Nat Rev Mol Cell Bio"},{"issue":"10","key":"pone.0251411.ref002","doi-asserted-by":"crossref","first-page":"e46107","DOI":"10.1371\/journal.pone.0046107","article-title":"Fabrication of hydrogels with steep stiffness gradients for studying cell mechanical response","volume":"7","author":"R Sunyer","year":"2012","journal-title":"PLoS ONE"},{"issue":"9","key":"pone.0251411.ref003","doi-asserted-by":"crossref","first-page":"733","DOI":"10.1038\/nmeth.1487","article-title":"Mechanical regulation of cell function with geometrically modulated elastomeric substrates","volume":"7","author":"J Fu","year":"2010","journal-title":"Nat Methods"},{"key":"pone.0251411.ref004","doi-asserted-by":"crossref","first-page":"757","DOI":"10.1038\/nnano.2012.167","article-title":"The nanomechanical signature of breast cancer","volume":"7","author":"M Plodinec","year":"2012","journal-title":"Nat Nanotech"},{"issue":"4","key":"pone.0251411.ref005","doi-asserted-by":"crossref","first-page":"265","DOI":"10.1038\/nrm1890","article-title":"Local force and geometry sensing regulate cell functions","volume":"7","author":"V Vogel","year":"2006","journal-title":"Nat Rev Mol Cell Bio"},{"issue":"22","key":"pone.0251411.ref006","doi-asserted-by":"crossref","first-page":"4605","DOI":"10.1242\/jcs.00792","article-title":"Early molecular events in the assembly of matrix adhesions at the leading edge of migrating cells","volume":"116","author":"R Zaidel-Bar","year":"2003","journal-title":"J Cell Sci"},{"issue":"1","key":"pone.0251411.ref007","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1002\/jcp.20550","article-title":"Mechanical forces alter zyxin unbinding kinetics within focal adhesions of living cells","volume":"207","author":"TP Lele","year":"2006","journal-title":"J Cell Physiol"},{"issue":"Pt 10","key":"pone.0251411.ref008","doi-asserted-by":"crossref","first-page":"1665","DOI":"10.1242\/jcs.042986","article-title":"Mechanosensing in actin stress fibers revealed by a close correlation between force and protein localization","volume":"122","author":"J Colombelli","year":"2009","journal-title":"J Cell Sci"},{"issue":"1","key":"pone.0251411.ref009","doi-asserted-by":"crossref","first-page":"9788","DOI":"10.1038\/s41598-018-27948-3","article-title":"Live cell imaging reveals focal adhesions mechanoresponses in mammary epithelial cells under sustained equibiaxial stress","volume":"8","author":"L Sigaut","year":"2018","journal-title":"Sci Rep"},{"issue":"46","key":"pone.0251411.ref010","first-page":"2173","article-title":"Preparation of complaint matrices for quantifying cellular contraction","author":"Y Aratyn-Schaus","year":"2010","journal-title":"JoVE"},{"issue":"2","key":"pone.0251411.ref011","doi-asserted-by":"crossref","first-page":"79","DOI":"10.1016\/S0962-8924(01)02205-X","article-title":"Flexible substrata for the detection of cellular traction forces","volume":"12","author":"KA Beningo","year":"2002","journal-title":"Trends Cell Biol"},{"issue":"1","key":"pone.0251411.ref012","doi-asserted-by":"crossref","first-page":"68","DOI":"10.1007\/s12195-010-0102-6","article-title":"Substrate Stiffness and Cell Area Predict Cellular Traction Stresses in Single Cells and Cells in Contact","volume":"3","author":"JP Califano","year":"2010","journal-title":"Cell Mol Bioeng"},{"issue":"4","key":"pone.0251411.ref013","doi-asserted-by":"crossref","first-page":"640","DOI":"10.1016\/j.bpj.2012.07.023","article-title":"Decoupling substrate stiffness, spread area, and micropost density: a close spatial relationship between traction forces and focal adhesions","volume":"103","author":"SJ Han","year":"2012","journal-title":"Biophys J"},{"issue":"11","key":"pone.0251411.ref014","doi-asserted-by":"crossref","first-page":"1454","DOI":"10.1039\/c5ib00217f","article-title":"Fibronectin fibrillogenesis facilitates mechano-dependent cell spreading, force generation, and nuclear size in human embryonic fibroblasts","volume":"7","author":"LE Scott","year":"2015","journal-title":"Integr Biol (Camb)"},{"issue":"2","key":"pone.0251411.ref015","doi-asserted-by":"crossref","first-page":"e32572","DOI":"10.1371\/journal.pone.0032572","article-title":"Cellular traction stresses increase with increasing metastatic potential","volume":"7","author":"CM Kraning-Rush","year":"2012","journal-title":"PLoS ONE"},{"issue":"1","key":"pone.0251411.ref016","doi-asserted-by":"crossref","first-page":"7228","DOI":"10.1038\/s41598-018-25589-0","article-title":"The mechanical microenvironment regulates ovarian cancer cell morphology, migration, and spheroid disaggregation","volume":"8","author":"AJ McKenzie","year":"2018","journal-title":"Sci Rep"},{"key":"pone.0251411.ref017","first-page":"521","volume-title":"Methods Cell Biol","author":"AJ Engler","year":"2007"},{"key":"pone.0251411.ref018","first-page":"1","volume-title":"An introduction to MatPIV v. 1.6.1., Department of Mathematics","author":"JK Sveen","year":"2004"},{"issue":"1","key":"pone.0251411.ref019","doi-asserted-by":"crossref","first-page":"207","DOI":"10.1529\/biophysj.107.113670","article-title":"High resolution traction force microscopy based on experimental and computational advances","volume":"94","author":"B Sabass","year":"2008","journal-title":"Biophys J"},{"key":"pone.0251411.ref020","first-page":"1035\/8","article-title":"Solution of Incorrectly Formulated Problems and the Regularization Method","volume":"5","author":"A Tikhonov","year":"1963","journal-title":"Soviet Math Dokl"},{"key":"pone.0251411.ref021","first-page":"119","article-title":"The L-curve and its use in the numerical treatment of inverse problems. Computational Inverse Problems in Electrocardiology","volume":"5","author":"P Hansen","year":"2001","journal-title":"Adv Comp Bioeng"},{"issue":"7","key":"pone.0251411.ref022","doi-asserted-by":"crossref","first-page":"2819","DOI":"10.1529\/biophysj.107.104984","article-title":"Paxillin dynamics measured during adhesion assembly and disassembly by correlation spectroscopy","volume":"94","author":"MA Digman","year":"2008","journal-title":"Biophys J"},{"issue":"2","key":"pone.0251411.ref023","doi-asserted-by":"crossref","first-page":"707","DOI":"10.1016\/j.bpj.2008.09.051","article-title":"Detecting protein complexes in living cells from laser scanning confocal image sequences by the cross correlation raster image spectroscopy method","volume":"96","author":"MA Digman","year":"2009","journal-title":"Biophys J"},{"key":"pone.0251411.ref024","doi-asserted-by":"crossref","first-page":"e02257","DOI":"10.7554\/eLife.02257","article-title":"Symmetric exchange of multi-protein building blocks between stationary focal adhesions and the cytosol","volume":"3","author":"J-E Hoffmann","year":"2014","journal-title":"eLife"},{"issue":"9","key":"pone.0251411.ref025","doi-asserted-by":"crossref","first-page":"1425","DOI":"10.1242\/jcs.077388","article-title":"Actomyosin-generated tension controls the molecular kinetics of focal adhesions","volume":"124","author":"H Wolfenson","year":"2011","journal-title":"J Cell Sci"},{"issue":"6","key":"pone.0251411.ref026","doi-asserted-by":"crossref","first-page":"1175","DOI":"10.1002\/jcb.20761","article-title":"Force meets chemistry: Analysis of mechanochemical conversion in focal adhesions using fluorescence recovery after photobleaching","volume":"97","author":"TP Lele","year":"2006","journal-title":"J Cell Biochem"},{"issue":"6","key":"pone.0251411.ref027","doi-asserted-by":"crossref","first-page":"3473","DOI":"10.1529\/biophysj.103.026765","article-title":"Analysis of Binding Reactions by Fluorescence Recovery after Photobleaching","volume":"86","author":"BL Sprague","year":"2004","journal-title":"Biophys J"},{"issue":"3","key":"pone.0251411.ref028","first-page":"181","article-title":"An improved mathematical approach for determination of molecular kinetics in living cells with FRAP","volume":"1","author":"T Lele","year":"2004","journal-title":"Mech Chem Biosyst"},{"issue":"14","key":"pone.0251411.ref029","doi-asserted-by":"crossref","first-page":"1901","DOI":"10.1091\/mbc.e17-02-0116","article-title":"Effects of substrate stiffness and actomyosin contractility on coupling between force transmission and vinculin-paxillin recruitment at single focal adhesions","volume":"28","author":"DW Zhou","year":"2017","journal-title":"Mol Biol Cell"},{"issue":"11, Part B","key":"pone.0251411.ref030","first-page":"3095","article-title":"Traction force microscopy on soft elastic substrates: A guide to recent computational advances","volume":"1853","author":"US Schwarz","year":"2015","journal-title":"BBA-Mol Cell Res"},{"issue":"7323","key":"pone.0251411.ref031","doi-asserted-by":"crossref","first-page":"580","DOI":"10.1038\/nature09621","article-title":"Nanoscale architecture of integrin-based cell adhesions","volume":"468","author":"P Kanchanawong","year":"2010","journal-title":"Nature"},{"issue":"24","key":"pone.0251411.ref032","doi-asserted-by":"crossref","first-page":"9788","DOI":"10.1073\/pnas.1216209110","article-title":"How vinculin regulates force transmission","volume":"110","author":"DW Dumbauld","year":"2013","journal-title":"Proc Natl Acad Sci U S A"},{"issue":"7","key":"pone.0251411.ref033","doi-asserted-by":"crossref","first-page":"1868","DOI":"10.1063\/1.1143970","article-title":"Calibration of atomic\u2010force microscope tips","volume":"64","author":"JL Hutter","year":"1993","journal-title":"Rev Sci Instrum"},{"issue":"3","key":"pone.0251411.ref034","doi-asserted-by":"crossref","first-page":"430","DOI":"10.1115\/1.2720924","article-title":"Robust Strategies for Automated AFM Force Curve Analysis\u2014I. Non-adhesive Indentation of Soft, Inhomogeneous Materials","volume":"129","author":"DC Lin","year":"2006","journal-title":"J Biomech Eng"},{"issue":"1","key":"pone.0251411.ref035","first-page":"47","article-title":"The relation between load and penetration in the axisymmetric boussinesq problem for a punch of arbitrary profile","volume":"3","author":"IN Sneddon","year":"1965","journal-title":"J Biomed Eng"},{"issue":"3","key":"pone.0251411.ref036","doi-asserted-by":"crossref","first-page":"235","DOI":"10.1080\/15583724.2010.493255","article-title":"Probing Soft Matter with the Atomic Force Microscopies: Imaging and Force Spectroscopy","volume":"50","author":"ME McConney","year":"2010","journal-title":"Polym Rev"},{"issue":"8","key":"pone.0251411.ref037","doi-asserted-by":"crossref","first-page":"4027","DOI":"10.1128\/MCB.10.8.4027","article-title":"Epidermal growth factor receptor, but not c-erbB-2, activation prevents lactogenic hormone induction of the beta-casein gene in mouse mammary epithelial cells","volume":"10","author":"NE Hynes","year":"1990","journal-title":"Mol Cell Biol"},{"key":"pone.0251411.ref038","doi-asserted-by":"crossref","first-page":"676","DOI":"10.1038\/nmeth.2019","article-title":"Fiji: an open-source platform for biological-image analysis","volume":"9","author":"J Schindelin","year":"2012","journal-title":"Nat Methods"},{"issue":"2","key":"pone.0251411.ref039","doi-asserted-by":"crossref","first-page":"365","DOI":"10.1016\/S0039-9140(01)00602-6","article-title":"Diffusion coefficient measurements in microfluidic devices","volume":"56","author":"CT Culbertson","year":"2002","journal-title":"Talanta"},{"issue":"4","key":"pone.0251411.ref040","doi-asserted-by":"crossref","first-page":"873","DOI":"10.1016\/S0006-3495(90)82432-2","article-title":"Quantification of transport and binding parameters using fluorescence recovery after photobleaching. Potential for in vivo applications","volume":"58","author":"EN Kaufman","year":"1990","journal-title":"Biophys J"}],"container-title":["PLOS ONE"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0251411","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2021,5,11]],"date-time":"2021-05-11T17:27:49Z","timestamp":1620754069000},"score":1,"resource":{"primary":{"URL":"https:\/\/dx.plos.org\/10.1371\/journal.pone.0251411"}},"subtitle":[],"editor":[{"given":"Colin","family":"Johnson","sequence":"first","affiliation":[],"role":[{"role":"editor","vocabulary":"crossref"}]}],"short-title":[],"issued":{"date-parts":[[2021,5,11]]},"references-count":40,"journal-issue":{"issue":"5","published-online":{"date-parts":[[2021,5,11]]}},"URL":"https:\/\/doi.org\/10.1371\/journal.pone.0251411","relation":{},"ISSN":["1932-6203"],"issn-type":[{"value":"1932-6203","type":"electronic"}],"subject":[],"published":{"date-parts":[[2021,5,11]]}}}