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Natl. Acad. Sci. U.S.A."],"published-print":{"date-parts":[[2026,3,31]]},"abstract":"<jats:p>Clathrin-mediated endocytosis (CME) is a multistage process that involves the initiation and stabilization of clathrin-coated pits (CCPs) that invaginate and finally detach from the plasma membrane to form clathrin-coated vesicles (CCVs). Given that Soluble N-ethylmaleimide-sensitive factor attachment receptor (SNARE) proteins are essential for downstream vesicle targeting and fusion events, their recruitment into nascent CCVs has been suggested to be a prerequisite for CME progression. However, which and how SNARE proteins regulate CME remains to be explored. Here, we showed that siRNA-mediated knockdown of the R-SNARE, vesicle-associated membrane protein 8 (VAMP8) impairs CCP initiation, stabilization, and invagination and strongly inhibits CME. Mechanistically, recruitment of VAMP8 to CCVs is not required for CME. Instead, depletion of VAMP8 inhibits recycling of endocytic cargoes and as exemplified here by transferrin receptor, skews their trafficking toward lysosomal degradation. VAMP8 depletion therefore indirectly impairs CCV formation and inhibits CME by depleting endocytic cargo. Overall, our study provides insights into the crosstalk between endocytosis and endocytic recycling of CME cargo and demonstrates the critical role for cargo recruitment in stabilizing nascent CCPs to regulate CME.<\/jats:p>","DOI":"10.1073\/pnas.2602991123","type":"journal-article","created":{"date-parts":[[2026,3,26]],"date-time":"2026-03-26T17:17:27Z","timestamp":1774545447000},"update-policy":"https:\/\/doi.org\/10.1073\/pnas.cm10313","source":"Crossref","is-referenced-by-count":0,"title":["VAMP8 function reveals tight linkage between endocytic recycling and endocytosis"],"prefix":"10.1073","volume":"123","author":[{"ORCID":"https:\/\/orcid.org\/0009-0007-5932-7728","authenticated-orcid":false,"given":"Ailing","family":"Liu","sequence":"first","affiliation":[{"name":"Department of Clinical Laboratory of The First Affiliated Hospital, Ministry of Education Key Laboratory of Rare Pediatric Diseases, Institute of Cytology and Genetics, Hengyang Medical School, University of South 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Aguet cmeAnalysis. GitHub. https:\/\/github.com\/DanuserLab\/cmeAnalysis. Deposited 29 April 2020."},{"key":"e_1_3_4_68_2","unstructured":"A. Liu VAMP8 function reveals tight linkage between endocyticrecycling and endocytosis. Figshare. https:\/\/figshare.com\/s\/e965e7ad44de55d2b283. Deposited 2 March 2026."},{"key":"e_1_3_4_69_2","unstructured":"A. Liu VAMP8 function reveals tight linkage between endocyticrecycling and endocytosis. iProX. https:\/\/www.iprox.cn\/page\/project.html?id=IPX0016058000. 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