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We fluorescently labeled the SARS\u2010CoV\u20102 RBD and its receptor ACE2 protein with small molecule dyes. Utilizing SIM imaging, the real\u2010time impact of inhibitors (BafA1, CQ, HCQ, Dynasore) on the RBD\u2010ACE2 endocytotic process was dynamically tracked in living cells. Initially, the protein activity of RBD and ACE2 was ensured after being labeled. And then our findings revealed that these inhibitors could inhibit the internalization and degradation of RBD\u2010ACE2 to varying degrees. Among them, 100\u2005nM BafA1 exhibited the most satisfactory endocytotic inhibition (~63.9\u2009%) and protein degradation inhibition (~97.7\u2009%). And it could inhibit the fusion between endocytic vesicles in the living cells. Additionally, Dynasore, a widely recognized dynein inhibitor, also demonstrated cell acidification inhibition effects. Together, these inhibitors collectively hinder SARS\u2010CoV\u20102 infection by inhibiting both the viral internalization and RNA release. The comprehensive evaluation of pharmacological mechanisms through super\u2010resolution fluorescence imaging has laid a crucial theoretical foundation for the development of potential drugs to treat COVID\u201019.<\/jats:p>","DOI":"10.1002\/cbic.202400404","type":"journal-article","created":{"date-parts":[[2024,6,14]],"date-time":"2024-06-14T23:10:25Z","timestamp":1718406625000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Super\u2010Resolution Imaging Reveals the Mechanism of Endosomal Acidification Inhibitors Against SARS\u2010CoV\u20102 Infection"],"prefix":"10.1002","volume":"25","author":[{"given":"Chunyu","family":"Yan","sequence":"first","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"},{"name":"School of Chemistry Dalian University of Technology  2 Linggong Road Dalian 116024 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Wei","family":"Zhou","sequence":"additional","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"},{"name":"School of Chemistry Dalian University of Technology  2 Linggong Road Dalian 116024 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xuelian","family":"Zhou","sequence":"additional","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"},{"name":"School of Chemistry Dalian University of Technology  2 Linggong Road Dalian 116024 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Qinglong","family":"Qiao","sequence":"additional","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Lu","family":"Miao","sequence":"additional","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-2491-8938","authenticated-orcid":false,"given":"Zhaochao","family":"Xu","sequence":"additional","affiliation":[{"name":"Dalian Institute of Chemical Physics, Chinese Academy of Sciences  457 Zhongshan Road Dalian 116023 China"},{"name":"School of Chemistry Dalian University of Technology  2 Linggong Road Dalian 116024 China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2024,7,29]]},"reference":[{"key":"e_1_2_9_1_1","unstructured":"\u00a0"},{"key":"e_1_2_9_2_2","doi-asserted-by":"publisher","DOI":"10.3390\/vaccines9040349"},{"key":"e_1_2_9_3_2","doi-asserted-by":"publisher","DOI":"10.1055\/s-0042-1759684"},{"key":"e_1_2_9_4_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41392-020-00243-2"},{"key":"e_1_2_9_5_1","doi-asserted-by":"crossref","unstructured":"B. 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