{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,24]],"date-time":"2026-09-24T16:19:01Z","timestamp":1790266741357,"version":"4.1.0"},"reference-count":34,"publisher":"American Chemical Society (ACS)","issue":"26","license":[{"start":{"date-parts":[[2020,6,23]],"date-time":"2020-06-23T00:00:00Z","timestamp":1592870400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/pubs.acs.org\/page\/policy\/authorchoice_termsofuse.html"}],"funder":[{"DOI":"10.13039\/501100002338","name":"Ministry of Education of the People's Republic of China","doi-asserted-by":"publisher","award":["21616326"],"award-info":[{"award-number":["21616326"]}],"id":[{"id":"10.13039\/501100002338","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2016M602606"],"award-info":[{"award-number":["2016M602606"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100002858","name":"China Postdoctoral Science Foundation","doi-asserted-by":"publisher","award":["2017T100661"],"award-info":[{"award-number":["2017T100661"]}],"id":[{"id":"10.13039\/501100002858","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006469","name":"Fundo para o Desenvolvimento das Ci?ncias e da Tecnologia","doi-asserted-by":"publisher","award":["0029\/2019\/A"],"award-info":[{"award-number":["0029\/2019\/A"]}],"id":[{"id":"10.13039\/501100006469","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100006469","name":"Fundo para o Desenvolvimento das Ci?ncias e da Tecnologia","doi-asserted-by":"publisher","award":["111\/2015\/A"],"award-info":[{"award-number":["111\/2015\/A"]}],"id":[{"id":"10.13039\/501100006469","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81602360"],"award-info":[{"award-number":["81602360"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81672224"],"award-info":[{"award-number":["81672224"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81871809"],"award-info":[{"award-number":["81871809"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003453","name":"Natural Science Foundation of Guangdong Province","doi-asserted-by":"publisher","award":["2017A030313556"],"award-info":[{"award-number":["2017A030313556"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003453","name":"Natural Science Foundation of Guangdong Province","doi-asserted-by":"publisher","award":["2017A030313665"],"award-info":[{"award-number":["2017A030313665"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003453","name":"Natural Science Foundation of Guangdong Province","doi-asserted-by":"publisher","award":["2018A0303130103"],"award-info":[{"award-number":["2018A0303130103"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100003453","name":"Natural Science Foundation of Guangdong Province","doi-asserted-by":"publisher","award":["2019A1515011082"],"award-info":[{"award-number":["2019A1515011082"]}],"id":[{"id":"10.13039\/501100003453","id-type":"DOI","asserted-by":"publisher"}]},{"name":"Science and Technology Planning Project of Guangzhou","award":["201707010493"],"award-info":[{"award-number":["201707010493"]}]},{"name":"Medical Scientific Research Foundation, Guangdong Province","award":["A2016017"],"award-info":[{"award-number":["A2016017"]}]},{"name":"Tianhe District","award":["2018YZ001"],"award-info":[{"award-number":["2018YZ001"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":[],"published-print":{"date-parts":[[2020,7,7]]},"abstract":"<jats:title>Abstract<\/jats:title>\n                  <jats:p>Glucose metabolism is an essential process for energy production and cell survival for both normal and abnormal cellular metabolism. Several glucose transporter\/solute carrier 2A (GLUT\/SLC2A) superfamily members, including glucose transporter 1 (GLUT1), have been shown to mediate the cellular uptake of glucose in diverse cell types. GLUT1-mediated glucose uptake is a transient and rapid process; thus, the real-time monitoring of GLUT1 trafficking is pivotal for a better understanding of GLUT1 expression and GLUT1-dependent glucose uptake. In the present study, we established a rapid and effective method to visualize the trafficking of GLUT1 between the plasma membrane (PM) and endolysosomal system in live cells using an mCherry-EGFP-GLUT1 tandem fluorescence tracing system. We found that GLUT1 localized at the PM exhibited both red (mCherry) and green (EGFP) fluorescence (yellow when overlapping). However, a significant increase in red punctate fluorescence (mCherry is resistant to acidic pH), but not green fluorescence (EGFP is quenched by acidic pH), was observed upon glucose deprivation, indicating that the mCherry-EGFP-GLUT1 functional protein was trafficked to the acidic endolysosomal system. Besides, we were able to calculate the relative ratio of mCherry to EGFP by quantification of the translocation coefficient, which can be used as a readout for GLUT1 internalization and subsequent lysosomal degradation. Two mutants, mCherry-EGFP-GLUT1-S226D and mCherry-EGFP-GLUT1-\u0394C4, were also constructed, which indirectly confirmed the specificity of mCherry-EGFP-GLUT1 for monitoring GLUT1 trafficking. By using a series of endosomal (Rab5, Rab7, and Rab11) and lysosomal markers, we were able to define a model of GLUT1 trafficking in live cells in which upon glucose deprivation, GLUT1 dissociates from the PM and experiences a pH gradient from 6.8\u20136.1 in the early endosomes to 6.0\u20134.8 in the late endosomes and finally pH 4.5 in lysosomes, which is appropriate for degradation. In addition, our proof-of-concept study indicated that the pmCherry-EGFP-GLUT1 tracing system can accurately reflect endogenous changes in GLUT1 in response to treatment with the small molecule, andrographolide. Since targeting GLUT1 expression and GLUT1-dependent glucose metabolism is a promising therapeutic strategy for diverse types of cancers and certain other glucose addiction diseases, our study herein indicates that pmCherry-EGFP-GLUT1 can be utilized as a biosensor for GLUT1-dependent functional studies and potential small molecule screening.<\/jats:p>","DOI":"10.1021\/acsomega.0c01054","type":"journal-article","created":{"date-parts":[[2020,6,23]],"date-time":"2020-06-23T09:15:37Z","timestamp":1592903737000},"page":"15911-15921","source":"Crossref","is-referenced-by-count":29,"title":["Visualization of GLUT1 Trafficking\nin Live Cancer Cells by the Use of a Dual-Fluorescence Reporter"],"prefix":"10.1021","volume":"5","author":[{"given":"Zhen-Yan","family":"Li","sequence":"first","affiliation":[{"name":"Jinan University , , , ,","place":["Guangzhou, Guangdong, China, 510630"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Yu-Ling","family":"Shi","sequence":"additional","affiliation":[{"name":"Guangzhou University of Chinese Medicine , , , ,","place":["Guangzhou, Guangdong, China, 510405"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Guo-Xiong","family":"Liang","sequence":"additional","affiliation":[{"name":"Zhongshan Hospital of Traditional Chinese Medicine , , , ,","place":["Zhongshan, Guangdong, China, 528400"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie","family":"Yang","sequence":"additional","affiliation":[{"name":"Jinan University , , , ,","place":["Guangzhou, Guangdong, China, 510630"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Song-Kuan","family":"Zhuang","sequence":"additional","affiliation":[{"name":"Xiamen University , , , ,","place":["Xiamen, Fujian, China, 361102"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jie-Bin","family":"Lin","sequence":"additional","affiliation":[{"name":"Guangzhou University of Chinese Medicine , , , ,","place":["Guangzhou, Guangdong, China, 510405"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Abdelmoumin","family":"Ghodbane","sequence":"additional","affiliation":[{"name":"Jinan University , , , ,","place":["Guangzhou, Guangdong, China, 510630"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Man-Seng","family":"Tam","sequence":"additional","affiliation":[{"name":"IAN WO Medical Center , , ,","place":["Macao Special Administrative Region, Macao, China, 999078"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zu-Jian","family":"Liang","sequence":"additional","affiliation":[{"name":"Guangzhou University of Chinese Medicine , , , ,","place":["Guangzhou, Guangdong, China, 510405"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Zhen-Gang","family":"Zha","sequence":"additional","affiliation":[{"name":"Jinan University , , , ,","place":["Guangzhou, Guangdong, China, 510630"]}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7391-436X","authenticated-orcid":true,"given":"Huan-Tian","family":"Zhang","sequence":"additional","affiliation":[{"name":"Jinan University , , , ,","place":["Guangzhou, Guangdong, China, 510630"]}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"316","published-online":{"date-parts":[[2020,6,23]]},"reference":[{"key":"2026091511101170100_cit1","doi-asserted-by":"publisher","first-page":"660","DOI":"10.1002\/iub.375","article-title":"Amino acid ingestion\nand glucose metabolism--a review","volume":"62","author":"Gannon","year":"2010","journal-title":"IUBMB Life"},{"key":"2026091511101170100_cit2","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1016\/j.mam.2012.07.001","article-title":"The SLC2 (GLUT) family\nof membrane transporters","volume":"34","author":"Mueckler","year":"2013","journal-title":"Mol. Aspects Med."},{"key":"2026091511101170100_cit3","doi-asserted-by":"publisher","first-page":"654","DOI":"10.1002\/jcp.20166","article-title":"Molecular and cellular regulation\nof glucose transporter (GLUT) proteins in cancer","volume":"202","author":"Macheda","year":"2005","journal-title":"J. Cell. Physiol."},{"key":"2026091511101170100_cit4","doi-asserted-by":"publisher","first-page":"90","DOI":"10.1016\/j.yebeh.2018.06.010","article-title":"The glucose transporter type 1 (Glut1)\nsyndromes","volume":"91","author":"Koch","year":"2019","journal-title":"Epilepsy Behav."},{"key":"2026091511101170100_cit5","doi-asserted-by":"publisher","first-page":"43356","DOI":"10.18632\/oncotarget.17445","article-title":"The prognostic value\nof GLUT1 in cancers: a systematic review and meta-analysis","volume":"8","author":"Yu","year":"2017","journal-title":"Oncotarget"},{"key":"2026091511101170100_cit6","doi-asserted-by":"publisher","first-page":"221","DOI":"10.2174\/1389200216666150602145145","article-title":"Glucose Addiction in Cancer Therapy:\nAdvances and Drawbacks","volume":"16","author":"Granja","year":"2015","journal-title":"Curr. Drug Metab."},{"key":"2026091511101170100_cit7","doi-asserted-by":"publisher","first-page":"1556","DOI":"10.1002\/jcb.25817","article-title":"PPAR\u03b1\nPromotes Cancer Cell Glut1 Transcription Repression","volume":"118","author":"You","year":"2017","journal-title":"J. Cell. Biochem."},{"key":"2026091511101170100_cit8","doi-asserted-by":"publisher","first-page":"33367","DOI":"10.1074\/jbc.272.52.33367","article-title":"Transcriptional activation\nof the Glut1 gene in response to oxidative stress in L6 myotubes","volume":"272","author":"Kozlovsky","year":"1997","journal-title":"J. Biol. Chem."},{"key":"2026091511101170100_cit9","doi-asserted-by":"publisher","first-page":"84","DOI":"10.1016\/j.molcel.2017.05.020","article-title":"Autophagy-Dependent Shuttling of\nTBC1D5 Controls Plasma Membrane Translocation of GLUT1 and Glucose\nUptake","volume":"67","author":"Roy","year":"2017","journal-title":"Mol. 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Biol."},{"key":"2026091511101170100_cit15","doi-asserted-by":"publisher","first-page":"121","DOI":"10.1038\/nature13306","article-title":"Crystal structure of the human glucose transporter GLUT1","volume":"510","author":"Deng","year":"2014","journal-title":"Nature"},{"key":"2026091511101170100_cit16","doi-asserted-by":"publisher","first-page":"15523","DOI":"10.1021\/bi00047a018","article-title":"Kinetic\nanalysis of glucose transporter trafficking in fibroblasts and adipocytes","volume":"34","author":"Yeh","year":"1995","journal-title":"Biochemistry"},{"key":"2026091511101170100_cit17","doi-asserted-by":"publisher","first-page":"29130","DOI":"10.1038\/srep29130","article-title":"Determination of GLUT1 Oligomerization Parameters using\nBioluminescent Fo\u0308rster Resonance Energy Transfer","volume":"6","author":"Looyenga","year":"2016","journal-title":"Sci. Rep."},{"key":"2026091511101170100_cit18","doi-asserted-by":"publisher","first-page":"1167","DOI":"10.1016\/j.molcel.2013.01.035","article-title":"AMPK-dependent degradation of TXNIP upon energy\nstress leads to enhanced glucose uptake via GLUT1","volume":"49","author":"Wu","year":"2013","journal-title":"Mol. Cell"},{"key":"2026091511101170100_cit19","doi-asserted-by":"publisher","first-page":"998","DOI":"10.1038\/s41598-018-37367-z","article-title":"New insights into GluT1 mechanics\nduring glucose transfer","volume":"9","author":"Galochkina","year":"2019","journal-title":"Sci. Rep."},{"key":"2026091511101170100_cit20","doi-asserted-by":"publisher","first-page":"452","DOI":"10.4161\/auto.4451","article-title":"Dissection\nof the autophagosome maturation process by a novel reporter protein,\ntandem fluorescent-tagged LC3","volume":"3","author":"Kimura","year":"2007","journal-title":"Autophagy"},{"key":"2026091511101170100_cit21","doi-asserted-by":"publisher","first-page":"30387","DOI":"10.1074\/jbc.M113.490581","article-title":"Glucose-starved cells do not engage in prosurvival\nautophagy","volume":"288","author":"Rami\u0301rez-Peinado","year":"2013","journal-title":"J. Biol. Chem."},{"key":"2026091511101170100_cit22","doi-asserted-by":"publisher","first-page":"3511","DOI":"10.1042\/BCJ20170677","article-title":"Chemical biology probes of mammalian\nGLUT structure and function","volume":"475","author":"Holman","year":"2018","journal-title":"Biochem. J."},{"key":"2026091511101170100_cit23","doi-asserted-by":"publisher","first-page":"467","DOI":"10.1091\/mbc.11.2.467","article-title":"Rab7: a key\nto lysosome biogenesis","volume":"11","author":"Bucci","year":"2017","journal-title":"Mol. Biol. Cell"},{"key":"2026091511101170100_cit24","doi-asserted-by":"publisher","first-page":"234","DOI":"10.1016\/j.cell.2012.09.013","article-title":"SnapShot:\nMammalian Rab proteins in endocytic trafficking","volume":"151","author":"Galvez","year":"2012","journal-title":"Cell"},{"key":"2026091511101170100_cit25","doi-asserted-by":"publisher","first-page":"94ra70","DOI":"10.1126\/scitranslmed.3002394","article-title":"Targeting GLUT1 and the Warburg effect in renal cell carcinoma by\nchemical synthetic lethality","volume":"3","author":"Chan","year":"2011","journal-title":"Sci. Transl. Med."},{"key":"2026091511101170100_cit26","doi-asserted-by":"publisher","first-page":"2333","DOI":"10.4155\/fmc-2019-0052","article-title":"The progress and development\nof GLUT1 inhibitors targeting cancer energy metabolism","volume":"11","author":"Meng","year":"2019","journal-title":"Future Med. Chem."},{"key":"2026091511101170100_cit27","doi-asserted-by":"publisher","first-page":"45","DOI":"10.1002\/cbic.201900544","article-title":"Small-Molecule Inhibition\nof Glucose Transporters GLUT-1-4","volume":"21","author":"Reckzeh","year":"2019","journal-title":"ChemBioChem"},{"key":"2026091511101170100_cit28","doi-asserted-by":"publisher","first-page":"4575","DOI":"10.1002\/jcb.26122","article-title":"Andrographolide\nInduces Cell Cycle Arrest and Apoptosis of Chondrosarcoma by Targeting\nTCF-1\/SOX9 Axis","volume":"118","author":"Zhang","year":"2017","journal-title":"J. Cell. Biochem."},{"key":"2026091511101170100_cit29","doi-asserted-by":"publisher","first-page":"1890","DOI":"10.1016\/j.drudis.2019.05.017","article-title":"Advances and\nchallenges in developing andrographolide and its analogues as cancer\ntherapeutic agents","volume":"24","author":"Soo","year":"2019","journal-title":"Drug Discovery Today"},{"key":"2026091511101170100_cit30","first-page":"255","article-title":"Andrographolide\nand analogues in cancer prevention","volume":"7","author":"Mishra","year":"2015","journal-title":"Front. Biosci.,\nElite Ed."},{"key":"2026091511101170100_cit31","doi-asserted-by":"publisher","first-page":"529","DOI":"10.1007\/s00210-007-0240-0","article-title":"Mediation of beta-endorphin in andrographolide-induced\nplasma glucose-lowering action in type I diabetes-like animals","volume":"377","author":"Yu","year":"2008","journal-title":"Naunyn-Schmiedeberg\u2019s Arch. Pharmacol."},{"key":"2026091511101170100_cit32","doi-asserted-by":"publisher","first-page":"1042","DOI":"10.1016\/j.chembiol.2011.05.013","article-title":"A Sensitive and Quantitative Technique for Detecting\nAutophagic Events Based on Lysosomal Delivery","volume":"18","author":"Katayama","year":"2011","journal-title":"Chem. Biol."},{"key":"2026091511101170100_cit33","doi-asserted-by":"publisher","first-page":"2182","DOI":"10.1002\/jcb.25863","article-title":"The BET Bromodomain Inhibitor JQ1\nSuppresses Chondrosarcoma Cell Growth via Regulation of YAP\/p21\/c-Myc\nSignaling","volume":"118","author":"Zhang","year":"2017","journal-title":"J. Cell. Biochem."},{"key":"2026091511101170100_cit34","doi-asserted-by":"publisher","first-page":"3894","DOI":"10.1002\/jcp.29283","article-title":"HSP90 regulates osteosarcoma\ncell apoptosis by targeting the p53\/TCF-1-mediated transcriptional\nnetwork","volume":"235","author":"Yang","year":"2019","journal-title":"J. Cell. Physiol."}],"container-title":["ACS Omega"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/pubs.acs.org\/doi\/pdf\/10.1021\/acsomega.0c01054","content-type":"application\/pdf","content-version":"vor","intended-application":"unspecified"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/5\/26\/15911\/67675027\/acsomega.0c01054.pdf","content-type":"application\/pdf","content-version":"vor","intended-application":"syndication"},{"URL":"https:\/\/pubs.acs.org\/acsodf\/article-pdf\/5\/26\/15911\/67675027\/acsomega.0c01054.pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2026,9,15]],"date-time":"2026-09-15T15:51:32Z","timestamp":1789487492000},"score":1,"resource":{"primary":{"URL":"https:\/\/pubs.acs.org\/acsodf\/article\/5\/26\/15911\/1648006\/Visualization-of-GLUT1-Trafficking-in-Live-Cancer"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,6,23]]},"references-count":34,"journal-issue":{"issue":"26","published-online":{"date-parts":[[2020,6,23]]},"published-print":{"date-parts":[[2020,7,7]]}},"URL":"https:\/\/doi.org\/10.1021\/acsomega.0c01054","relation":{},"ISSN":["2470-1343"],"issn-type":[{"value":"2470-1343","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,6,23]]}}}