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Whether the toxin induces its own pathway or travels along a physiological retrograde route is not known. To study its trafficking, we labeled cholera toxin B (<jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content>) or endogenous plasma membrane proteins with a small chemical compound, benzylguanine, which covalently reacts with the protein <jats:styled-content style=\"fixed-case\">SNAP<\/jats:styled-content>\u2010tag. Using <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>\u2010targeted <jats:styled-content style=\"fixed-case\">SNAP<\/jats:styled-content>\u2010tag as reporter, we found that transport of <jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content> to the <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content> depends on dynamin\u20102 and syntaxin 5. Plasma membrane proteins and a fluid\u2010phase marker added to the medium were also transported to the <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>. This flux was not affected by exposing cells to <jats:styled-content style=\"fixed-case\">CTB<\/jats:styled-content> but was inhibited by depleting syntaxin 5 and increased by depleting dynamin\u20102. As a control for confined intracellular localization of <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>\u2010targeted <jats:styled-content style=\"fixed-case\">SNAP<\/jats:styled-content>\u2010tag we used adenovirus\u20105, which traffics to endosomes and then escapes into the cytosol. The virus did not react with <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content>\u2010targeted <jats:styled-content style=\"fixed-case\">SNAP<\/jats:styled-content> but with cytosolic <jats:styled-content style=\"fixed-case\">SNAP<\/jats:styled-content>. Together, our results establish a new method (<jats:styled-content style=\"fixed-case\">SNAP<\/jats:styled-content>\u2010trap) to study trafficking of different cargo to the <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content> and the cytosol and provide evidence for the existence of a constitutive pathway from the cell surface to the <jats:styled-content style=\"fixed-case\">ER<\/jats:styled-content><\/jats:bold>.<\/jats:p>","DOI":"10.1111\/tra.12018","type":"journal-article","created":{"date-parts":[[2012,10,9]],"date-time":"2012-10-09T18:04:31Z","timestamp":1349805871000},"page":"36-46","source":"Crossref","is-referenced-by-count":19,"title":["Investigating Endocytic Pathways to the Endoplasmic Reticulum and to the Cytosol Using <scp>SNAP<\/scp>\u2010Trap"],"prefix":"10.1111","volume":"14","author":[{"given":"Roger","family":"Geiger","sequence":"first","affiliation":[{"name":"Institute of Biochemistry ETH Zurich  Schafmattstrasse 18 Zurich CH\u20108093 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