Lipid droplets are the place for HCV assembly and ADRP is an abundant lipid droplets-associated protein. However, little is known about the mechanisms how ADRP is involved in HCV life cycle. Here we demonstrate that activation of ARF1 dissociates ADRP from lipid droplets. A constitute active form of ARF1 (ARF1Q71I) promotes HCV assembly. We found that ADRP plays a positive role in HCV replication and a negative role in HCV assembly. Overexpression of ADRP increases the size of lipid droplets, while silencing ADRP reduces the size of lipid droplets. These findings provide new insight into the role of lipid droplets proteins in life cycle of HCV. (C) 2016 Elsevier Inc. All rights reserved.