Claudin-1 is a HCV co-receptor required for viral entry.While extensive studies have focused on claudin-1 as an anti-HCV target,little is known about how claudin-1 is regulated by host vesicular transport.Here we found that claudin-1 associated with Sec24C,a cargo sorting component of COPⅡ.Sec24C was partially colocalized with claudin-1.The carboxyl terminal YV motif in claudin-1 was required for the interaction between claudin-1 and Sec24C,tight junction localization of claudin-1,and HCV entry.Blocking COPⅡ transport by siRNA or inhibitor reduced surface level of claudin-1 and HCV entry.Moreover,inhibition of HCV entry by siRNA against claudin-1 was rescued by siRNA-resistant claudin-1 but not claudin-1-YV/AA.Our data identified claudin-1 as a cargo of COPⅡ transport.By interacting with the cargo sorting component of the COPⅡ machinery,claudin-1 is transported from ER to Golgi,eventually to tight junction.Blocking COPⅡ transport will inhibit HCV entry through reducing protein level of claudin-1 in tight junction.Our work will shed mechanistic insight on how HCV entry is regulated by COPⅡ transport.