{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,12]],"date-time":"2025-10-12T04:37:27Z","timestamp":1760243847770,"version":"build-2065373602"},"reference-count":41,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2011,2,10]],"date-time":"2011-02-10T00:00:00Z","timestamp":1297296000000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>The glucose transporter 4 (GLUT4) plays a key role in maintaining whole body glucose homeostasis. Tracking GLUT4 in space and time can provide new insights for understanding the mechanisms of insulin-regulated GLUT4 translocation. Organic dyes and fluorescent proteins were used in previous studies for investigating the traffic of GLUT4 in skeletal muscle cells and adipocytes. Because of their relative weak fluorescent signal against strong cellular autofluorescence background and their fast photobleaching rate, most studies only focused on particular segments of GLUT4 traffic. In this study, we have developed a new method for observing the translocation of GLUT4 targeted with photostable and bright quantum dots (QDs) in live L6 cells. QDs were targeted to GLUT4myc specifically and internalized with GLUT4myc through receptor-mediated endocytosis. Compared with traditional fluorescence dyes and fluorescent proteins, QDs with high brightness and extremely photostability are suitable for long-term single particle tracking, so individual GLUT4-QD complex can be easily detected and tracked for long periods of time. This newly described method will be a powerful tool for observing the translocation of GLUT4 in live L6 cells.<\/jats:p>","DOI":"10.3390\/s110202077","type":"journal-article","created":{"date-parts":[[2011,2,10]],"date-time":"2011-02-10T14:52:26Z","timestamp":1297349546000},"page":"2077-2089","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Observing GLUT4 Translocation in Live L6 Cells Using Quantum Dots"],"prefix":"10.3390","volume":"11","author":[{"given":"Feng","family":"Qu","sequence":"first","affiliation":[{"name":"Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zubin","family":"Chen","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiaoxuan","family":"Wang","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lingfeng","family":"Meng","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Zhengxing","family":"Wu","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Anlian","family":"Qu","sequence":"additional","affiliation":[{"name":"Institute of Biophysics and Biochemistry, School of Life Science and Technology, Huazhong University of Science and Technology, Wuhan 430074, China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2011,2,10]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"248","DOI":"10.1056\/NEJM199907223410406","article-title":"Glucose transporters and insulin action\u2014implications for insulin resistance and diabetes mellitus","volume":"341","author":"Shepherd","year":"1999","journal-title":"N. Engl. J. Med"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1038\/nrm782","article-title":"Regulated transport of the glucose transporter GLUT4","volume":"3","author":"Bryant","year":"2002","journal-title":"Nat. Rev. Mol. Cell Biol"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1016\/j.cellsig.2006.05.018","article-title":"How many signals impinge on GLUT4 activation by insulin?","volume":"19","author":"He","year":"2007","journal-title":"Cell Signal"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"1568","DOI":"10.1172\/JCI116025","article-title":"Insulin resistance in obese Zucker rat (fa\/fa) skeletal muscle is associated with a failure of glucose transporter translocation","volume":"90","author":"King","year":"1992","journal-title":"J. Clin. Invest"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"1180","DOI":"10.1007\/BF02658504","article-title":"Insulin action on glucose transport and plasma membrane GLUT4 content in skeletal muscle from patients with NIDDM","volume":"39","author":"Zierath","year":"1996","journal-title":"Diabetologia"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"821","DOI":"10.1007\/s001250051457","article-title":"Insulin action and insulin resistance in human skeletal muscle","volume":"43","author":"Zierath","year":"2000","journal-title":"Diabetologia"},{"key":"ref_7","first-page":"271","article-title":"Turning signals on and off: GLUT4 traffic in the insulin-signaling highway","volume":"20","author":"Thong","year":"2005","journal-title":"Physiology (Bethesda)"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"297","DOI":"10.1046\/j.1365-201X.2003.01163.x","article-title":"Push\/pull mechanisms of GLUT4 traffic in muscle cells","volume":"178","author":"Rudich","year":"2003","journal-title":"Acta Physiol. Scand"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"44212","DOI":"10.1074\/jbc.M102964200","article-title":"Insulin accelerates inter-endosomal GLUT4 traffic via phosphatidylinositol 3-kinase and protein kinase B","volume":"276","author":"Foster","year":"2001","journal-title":"J. Biol. Chem"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"1137","DOI":"10.1038\/sj.embor.7400584","article-title":"Glucose transporter 4: Cycling, compartments and controversies","volume":"6","author":"Dugani","year":"2005","journal-title":"EMBO Rep"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"193","DOI":"10.1016\/S0014-5793(98)00423-2","article-title":"GLUT4 translocation by insulin in intact muscle cells: Detection by a fast and quantitative assay","volume":"427","author":"Wang","year":"1998","journal-title":"FEBS Lett"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"4065","DOI":"10.1242\/jcs.113.22.4065","article-title":"Demonstration of insulin-responsive trafficking of GLUT4 and vpTR in fibroblasts","volume":"113","author":"Lampson","year":"2000","journal-title":"J. Cell Sci"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"445","DOI":"10.1006\/bbrc.2001.5620","article-title":"Insulin-regulated trafficking of dual-labeled glucose transporter 4 in primary rat adipose cells*1","volume":"287","author":"Dawson","year":"2001","journal-title":"Biochem. Biophys. Res. Commun"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1016\/j.cmet.2006.11.013","article-title":"Dissecting multiple steps of GLUT4 trafficking and identifying the sites of insulin action","volume":"5","author":"Bai","year":"2007","journal-title":"Cell Metab"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/S0958-1669(02)00282-3","article-title":"Luminescent quantum dots for multiplexed biological detection and imaging","volume":"13","author":"Chan","year":"2002","journal-title":"Curr. Opin. Biotechnol"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"532","DOI":"10.1117\/1.1506706","article-title":"Quantum-dot nanocrystals for ultrasensitive biological labeling and multicolor optical encoding","volume":"7","author":"Gao","year":"2002","journal-title":"J. Biomed. Opt"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"496","DOI":"10.1016\/S0006-291X(03)00211-0","article-title":"Semiconductor quantum dot\/albumin complex is a long-life and highly photostable endosome marker","volume":"302","author":"Hanaki","year":"2003","journal-title":"Biochem. Biophys. Res. Commun"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"47","DOI":"10.1038\/nbt927","article-title":"The use of nanocrystals in biological detection","volume":"22","author":"Alivisatos","year":"2004","journal-title":"Nat. Biotechnol"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"63","DOI":"10.1016\/j.copbio.2004.11.003","article-title":"In vivo molecular and cellular imaging with quantum dots","volume":"16","author":"Gao","year":"2005","journal-title":"Curr. Opin. Biotechnol"},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Monton, H., Nogues, C., Rossinyol, E., Castell, O., and Roldan, M. (2009). QDs versus Alexa: reality of promising tools for immunocytochemistry. J. Nanobiotechnol.","DOI":"10.1186\/1477-3155-7-4"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"40","DOI":"10.1016\/S0958-1669(02)00282-3","article-title":"Luminescent quantum dots for multiplexed biological detection and imaging","volume":"13","author":"Chan","year":"2002","journal-title":"Curr. Opin. Biotechnol"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"198","DOI":"10.1038\/nbt929","article-title":"Quantum dot ligands provide new insights into erbB\/HER receptor-mediated signal transduction","volume":"22","author":"Lidke","year":"2004","journal-title":"Nat. Biotechnol"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1038\/nbt764","article-title":"Immunofluorescent labeling of cancer marker Her2 and other cellular targets with semiconductor quantum dots","volume":"21","author":"Wu","year":"2003","journal-title":"Nat. Biotechnol"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"841","DOI":"10.1021\/ac010662m","article-title":"Conjugation of luminescent quantum dots with antibodies using an engineered adaptor protein to provide new reagents for fluoroimmunoassays","volume":"74","author":"Goldman","year":"2002","journal-title":"Anal. Chem"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"825","DOI":"10.1364\/OL.26.000825","article-title":"Time-gated biological imaging by use of colloidal quantum dots","volume":"26","author":"Dahan","year":"2001","journal-title":"Opt Lett"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"73","DOI":"10.1038\/nmeth1004-73","article-title":"Use of quantum dots for live cell imaging","volume":"1","author":"Jaiswal","year":"2004","journal-title":"Nat. Methods"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"687","DOI":"10.1139\/O09-041","article-title":"Labeling and imaging of GLUT4 in live L6 cells with quantum dots","volume":"87","author":"Wang","year":"2009","journal-title":"Biochem. Cell Biol"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"228","DOI":"10.2337\/diacare.13.3.228","article-title":"Glucose transport and glucose transporters in muscle and their metabolic regulation","volume":"13","author":"Klip","year":"1990","journal-title":"Diabetes Care"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"2721","DOI":"10.1091\/mbc.e10-01-0029","article-title":"Identification of three distinct functional sites of insulin-mediated GLUT4 Trafficking in adipocytes using quantitative single molecule imaging","volume":"21","author":"Fujita","year":"2010","journal-title":"Mol. Biol. Cell"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"267","DOI":"10.1038\/nrm782","article-title":"Regulated transport of the glucose transporter GLUT4","volume":"3","author":"Bryant","year":"2002","journal-title":"Nat. Rev. Mol. Cell Biol"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"311","DOI":"10.1074\/jbc.M705756200","article-title":"Insulin releases Glut4 from static storage compartments into cycling endosomes and increases the rate constant for Glut4 exocytosis","volume":"283","author":"Muretta","year":"2008","journal-title":"J. Biol. Chem"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"2090","DOI":"10.2337\/diabetes.51.7.2090","article-title":"Sustained exposure of L6 myotubes to high glucose and insulin decreases insulin-stimulated GLUT4 translocation but upregulates GLUT4 activity","volume":"51","author":"Huang","year":"2002","journal-title":"Diabetes"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1002\/jbio.200810029","article-title":"Tracking bio-molecules in live cells using quantum dots","volume":"1","author":"Chang","year":"2008","journal-title":"J. Biophotonics"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"442","DOI":"10.1126\/science.1088525","article-title":"Diffusion dynamics of glycine receptors revealed by single-quantum dot tracking","volume":"302","author":"Dahan","year":"2003","journal-title":"Science"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"11251","DOI":"10.1073\/pnas.0702536104","article-title":"Asymmetric redistribution of GABA receptors during GABA gradient sensing by nerve growth cones analyzed by single quantum dot imaging","volume":"104","author":"Bouzigues","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"12433","DOI":"10.1523\/JNEUROSCI.3349-07.2007","article-title":"Surface trafficking of neurotransmitter receptor: Comparison between single-molecule\/quantum dot strategies","volume":"27","author":"Groc","year":"2007","journal-title":"J. Neurosci"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"13666","DOI":"10.1073\/pnas.0706192104","article-title":"One at a time, live tracking of NGF axonal transport using quantum dots","volume":"104","author":"Cui","year":"2007","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1827","DOI":"10.1021\/nl049170q","article-title":"Fluorescent CdSe\/ZnS nanocrystal-peptide conjugates for long-term, nontoxic imaging and nuclear targeting in living cells","volume":"4","author":"Chen","year":"2004","journal-title":"Nano Lett"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"961","DOI":"10.1002\/adma.200306111","article-title":"Intracellular delivery of quantum dots for live cell labeling and organelle tracking","volume":"16","author":"Derfus","year":"2004","journal-title":"Adv. Mater"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"985","DOI":"10.1111\/j.1348-0421.2004.tb03621.x","article-title":"Quantum dots targeted to the assigned organelle in living cells","volume":"48","author":"Hoshino","year":"2004","journal-title":"Microbiol. Immunol"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1343","DOI":"10.1039\/B618902D","article-title":"Bioanalytics and biolabeling with semiconductor nanoparticles (quantum dots)","volume":"17","author":"Lin","year":"2007","journal-title":"J. Mater. Chem"}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/11\/2\/2077\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T21:55:12Z","timestamp":1760219712000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/11\/2\/2077"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2011,2,10]]},"references-count":41,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2011,2]]}},"alternative-id":["s110202077"],"URL":"https:\/\/doi.org\/10.3390\/s110202077","relation":{},"ISSN":["1424-8220"],"issn-type":[{"type":"electronic","value":"1424-8220"}],"subject":[],"published":{"date-parts":[[2011,2,10]]}}}