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As endosomes mature to become multivesicular bodies (<jats:styled-content style=\"fixed-case\">MVBs<\/jats:styled-content>), Endosomal Sorting Complexes Required for Transport  (ESCRTs) selectively incorporate transmembrane cargos into vesicles that bud into the endosome lumen. Luminal vesicles and their cargoes are targeted for destruction when <jats:styled-content style=\"fixed-case\">MVBs<\/jats:styled-content> fuse with lysosomes. Common assays of endosomal luminal targeting, including fluorescence microscopy and monitoring of proteolytic cargo maturation, possess significant limitations. We present a quantitative assay system called LUCID (<jats:styled-content style=\"fixed-case\">LUCiferase<\/jats:styled-content> reporter of Intraluminal Deposition) that monitors exposure of chimeric luciferase\u2010cargo reporters to cytosol. Luciferase\u2010chimera signal increases when sorting to the endosome lumen is disrupted, and silencing of signal from the chimera depends upon luminal delivery of the reporter rather than proteolytic degradation. The system presents several advantages, including rapidity, microscale operation and a high degree of reproducibility that enables detection of subtle phenotypic differences. Luciferase reporters provide linear signal over an extremely broad dynamic range, allowing analysis of reporter traffic even at anemic levels of expression. Furthermore, <jats:styled-content style=\"fixed-case\">LUCID<\/jats:styled-content> reports transport kinetics when applied to inducible trafficking reporters.<\/jats:p><jats:p><jats:inline-graphic xmlns:xlink=\"http:\/\/www.w3.org\/1999\/xlink\" xlink:href=\"graphic\/tra12331-gra-0001.png\" xlink:title=\"image\"\/><\/jats:p>","DOI":"10.1111\/tra.12331","type":"journal-article","created":{"date-parts":[[2015,10,1]],"date-time":"2015-10-01T07:24:57Z","timestamp":1443684297000},"page":"1318-1329","source":"Crossref","is-referenced-by-count":8,"title":["<scp>LUCID<\/scp>: A Quantitative Assay of <scp>ESCRT<\/scp>\u2010Mediated Cargo Sorting into Multivesicular Bodies"],"prefix":"10.1111","volume":"16","author":[{"given":"Daniel P.","family":"Nickerson","sequence":"first","affiliation":[{"name":"Department of Biochemistry University of Washington  Seattle WA 98195\u20107350 USA"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alexey 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