{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,6]],"date-time":"2026-10-06T21:45:25Z","timestamp":1791323125544,"version":"4.3.1"},"reference-count":32,"publisher":"Oxford University Press (OUP)","issue":"3","license":[{"start":{"date-parts":[[2013,6,21]],"date-time":"2013-06-21T00:00:00Z","timestamp":1371772800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/academic.oup.com\/pages\/standard-publication-reuse-rights"}],"funder":[{"DOI":"10.13039\/501100001677","name":"INSERM","doi-asserted-by":"publisher","id":[{"id":"10.13039\/501100001677","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Minist\u00e8re de l'Education Nationale et de la Recherche"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2013,6,21]]},"abstract":"<jats:title>ABSTRACT<\/jats:title><jats:p>Actin cytoskeleton plays a critical role in regulating T cell motility and activation. However, the lack of a real-time quantitative method to analyze actin assembly has limited the progress toward understanding actin regulation. Here, we describe a novel approach to probe actin dynamics on living T cells using FRET combined with flow cytometry. We have first generated a Jurkat T cell line stably coexpressing EGFP and mOrange FPs fused to actin. The real-time variation of actin monomer assembly or disassembly into filaments was quantified using a ratiometric flow cytometry method measuring changes in the mOrange\/EGFP emission ratio. The method was validated on resting T cells by using chemical compounds with known effects on actin filaments and comparison with conventional microscopy imaging. Our method also detected the rapid and transient actin assembly in T cells stimulated by anti-CD3\/CD28-coated beads, demonstrating its robustness and high sensitivity. Finally, we provide evidence that lentiviral-mediated transduction of shRNAs in engineered Jurkat cells could be used as a strategy to identify regulators of actin remodeling. In conclusion, the flow cytometric FRET analysis of actin polymerization represents a new technical advance to study the dynamics of actin regulation in intact cells.<\/jats:p>","DOI":"10.1189\/jlb.0113022","type":"journal-article","created":{"date-parts":[[2013,6,22]],"date-time":"2013-06-22T01:24:14Z","timestamp":1371864254000},"page":"531-539","source":"Crossref","is-referenced-by-count":8,"title":["Technical Advance: Actin CytoFRET, a novel FRET flow cytometry method for detection of actin dynamics in resting and activated T cell"],"prefix":"10.1093","volume":"94","author":[{"given":"Fr\u00e9d\u00e9ric","family":"Larbret","sequence":"first","affiliation":[{"name":"Tol\u00e9rance Immunitaire, Universit\u00e9 de Nice Sophia-Antipolis , Nice, France"},{"name":"INSERM, U576, Flow Cytometry Facility, H\u00f4pital Archet 1 , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Nicholas","family":"Dubois","sequence":"additional","affiliation":[{"name":"INSERM, U1065, C3M, Microenvironnement, Signalisation et Cancer , Nice, France"},{"name":"Facult\u00e9 de M\u00e9decine, Universit\u00e9 de Nice Sophia-Antipolis , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fr\u00e9d\u00e9ric","family":"Brau","sequence":"additional","affiliation":[{"name":"CNRS, UMR6097, Institut de Pharmacologie Mol\u00e9culaire et Cellulaire , Valbonne, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Elodie","family":"Guillemot","sequence":"additional","affiliation":[{"name":"Facult\u00e9 de M\u00e9decine, Universit\u00e9 de Nice Sophia-Antipolis , Nice, France"},{"name":"FRE-CNRS 3472 , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Karim","family":"Mahiddine","sequence":"additional","affiliation":[{"name":"INSERM, U576, Flow Cytometry Facility, H\u00f4pital Archet 1 , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Sophie","family":"Tartare-Deckert","sequence":"additional","affiliation":[{"name":"INSERM, U1065, C3M, Microenvironnement, Signalisation et Cancer , Nice, France"},{"name":"Facult\u00e9 de M\u00e9decine, Universit\u00e9 de Nice Sophia-Antipolis , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Val\u00e9rie","family":"Verhasselt","sequence":"additional","affiliation":[{"name":"Tol\u00e9rance Immunitaire, Universit\u00e9 de Nice Sophia-Antipolis , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcel","family":"Deckert","sequence":"additional","affiliation":[{"name":"INSERM, U1065, C3M, Microenvironnement, Signalisation et Cancer , Nice, France"},{"name":"Facult\u00e9 de M\u00e9decine, Universit\u00e9 de Nice Sophia-Antipolis , Nice, France"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"286","published-online":{"date-parts":[[2013,6,21]]},"reference":[{"key":"2023032711582778300_","doi-asserted-by":"crossref","first-page":"30","DOI":"10.1189\/jlb.0602272","article-title":"Actin cytoskeletal dynamics in T lymphocyte activation and migration","volume":"73","author":"Samstag","year":"2003","journal-title":"J. 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