{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,5,27]],"date-time":"2026-05-27T07:04:57Z","timestamp":1779865497077,"version":"3.53.1"},"reference-count":55,"publisher":"Wiley","issue":"5","license":[{"start":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T00:00:00Z","timestamp":1778889600000},"content-version":"vor","delay-in-days":15,"URL":"http:\/\/creativecommons.org\/licenses\/by\/4.0\/"},{"start":{"date-parts":[[2026,5,1]],"date-time":"2026-05-01T00:00:00Z","timestamp":1777593600000},"content-version":"tdm","delay-in-days":0,"URL":"http:\/\/doi.wiley.com\/10.1002\/tdm_license_1.1"}],"funder":[{"DOI":"10.13039\/501100000925","name":"National Health and Medical Research Council","doi-asserted-by":"publisher","award":["APP1107109"],"award-info":[{"award-number":["APP1107109"]}],"id":[{"id":"10.13039\/501100000925","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100008436","name":"National Foundation for Medical Research and Innovation","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100008436","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Journal of Neurochemistry"],"published-print":{"date-parts":[[2026,5]]},"abstract":"<jats:title>ABSTRACT<\/jats:title>\n                  <jats:p>Amyloidogenic proteins, such as amyloid\u2010\u03b2 (A\u03b2), self\u2010assemble into cross\u2010\u03b2 fibrils whose accumulation is central to Alzheimer's disease (AD). Measuring A\u03b2 aggregation and clearance in living cells remains challenging using current cell\u2010based assays, which are often low\u2010throughput or not suited for real\u2010time monitoring. This study aimed to (1) develop a robust, quantitative, and scalable fluorescence\u2010based assay using Amytracker to monitor A\u03b2 accumulation and clearance in an A\u03b2\u2010producing neuronal cell model, and (2) validate its utility for mechanistic studies and therapeutic screening. We established a plate\u2010based fluorescence assay using Amytracker in MC65 neuronal AD model expressing the Amyloid precursor protein C\u2010terminal fragment (APP\u2010C99) that generates A\u03b2. Accumulation and clearance of A\u03b2 were quantified by measuring Amytracker fluorescence under basal conditions and after inducing A\u03b2 clearance using a Tet\u2010suppressible system. We utilized this assay to evaluate cell death inhibitors ferrostatin\u20101 and liproxstatin\u20101 and proteasome activator IU1. Specificity of the assay for amyloidogenic proteins was assessed by treating wild\u2010type neuroblastoma cells with A\u03b2, human islet amyloid polypeptide (hIAPP), or non\u2010aggregating A\u03b2 controls. Validation included A\u03b2 immunoblotting and cell viability assays. In APP\u2010C99 expressing cells, elevated Amytracker fluorescence correlated with increased A\u03b2 accumulation and reduced cell viability. Supplementation of ferrostatin\u20101, liproxstatin\u20101, and IU1, on these cells, markedly reduced Amytracker signal, indicating decreased A\u03b2 burden. Furthermore, the Amytracker assay specifically detected amyloidogenic protein aggregation: wild\u2010type cells exposed to A\u03b242 or hIAPP showed high fluorescence, whereas non\u2010aggregating A\u03b216 peptide did not. The Amytracker assay provides a simple, non\u2010toxic, and high\u2010throughput platform for quantifying A\u03b2 accumulation and clearance in live cell models. Its sensitivity, specificity, and compatibility with high\u2010throughput screening make it a valuable tool for studying A\u03b2 dynamics, interrogating mechanisms of proteostasis, and identifying therapeutic candidates targeting A\u03b2.<\/jats:p>\n                  <jats:p>\n                    <jats:boxed-text content-type=\"graphic\" position=\"anchor\">\n                      <jats:graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" mimetype=\"image\/png\" position=\"anchor\" specific-use=\"enlarged-web-image\" xlink:href=\"graphic\/jnc70473-gra-0001-m.png\">\n                        <jats:alt-text>image<\/jats:alt-text>\n                      <\/jats:graphic>\n                    <\/jats:boxed-text>\n                  <\/jats:p>","DOI":"10.1111\/jnc.70473","type":"journal-article","created":{"date-parts":[[2026,5,16]],"date-time":"2026-05-16T07:40:07Z","timestamp":1778917207000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":0,"title":["A High\u2010Throughput Assay for Monitoring and Quantifying Amyloid\u2010\u03b2 Accumulation and Clearance in Alzheimer's Disease Cell Models"],"prefix":"10.1111","volume":"170","author":[{"given":"Ajish","family":"Ariyath","sequence":"first","affiliation":[{"name":"Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Sarich and Patricia Neuroscience Research Institute Edith Cowan University  Joondalup Western Australia Australia"},{"name":"Alzheimer's Research Australia  Nedlands Western Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Fraulein Denise","family":"Arigo","sequence":"additional","affiliation":[{"name":"Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Sarich and Patricia Neuroscience Research Institute Edith Cowan University  Joondalup Western Australia Australia"},{"name":"Alzheimer's Research Australia  Nedlands Western Australia Australia"},{"name":"University of Western Australia Medical School  Perth Western Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Anna","family":"Fyfe","sequence":"additional","affiliation":[{"name":"Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Sarich and Patricia Neuroscience Research Institute Edith Cowan University  Joondalup Western Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8364-7808","authenticated-orcid":false,"given":"W. 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Binosha","family":"Fernando","sequence":"additional","affiliation":[{"name":"Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Sarich and Patricia Neuroscience Research Institute Edith Cowan University  Joondalup Western Australia Australia"},{"name":"Alzheimer's Research Australia  Nedlands Western Australia Australia"},{"name":"University of Western Australia Medical School  Perth Western Australia Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ralph","family":"Martins","sequence":"additional","affiliation":[{"name":"Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Sarich and Patricia Neuroscience Research Institute Edith Cowan University  Joondalup Western Australia Australia"},{"name":"Alzheimer's Research Australia  Nedlands Western Australia Australia"},{"name":"University of Western Australia Medical School  Perth Western Australia Australia"},{"name":"School of Biomedical Science Macquarie University  Sydney New South Wales Australia"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-4361-9906","authenticated-orcid":false,"given":"Prashant","family":"Bharadwaj","sequence":"additional","affiliation":[{"name":"Centre of Excellence for Alzheimer's Disease Research and Care, School of Medical and Health Sciences, Sarich and Patricia Neuroscience Research Institute Edith Cowan University  Joondalup Western Australia Australia"},{"name":"Alzheimer's Research Australia  Nedlands Western Australia Australia"},{"name":"University of Western Australia Medical School  Perth Western Australia Australia"},{"name":"Curtin Medical School, Curtin Health and Innovation Research Institute (CHIRI), Faculty of Health Sciences Curtin University  Western Australia 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