Figure 3
HBV capsid formation was attenuated in the cells treated with nocodazole. (A) HepG2 cells transfected with a reporter plasmid carrying the HBV core promoter upstream of the luciferase gene were treated with 10, 20 and 40 μM nocodazole or 30 μM HX531 as a positive control for 7 h, and luciferase activity driven by the HBV core promoter was measured. (B,C) HepG2 cells transfected with an HBV-encoding plasmid were treated with or without 1 μM Bay41–4109 or 1, 2.5, 5 and 10 μM nocodazole, and then total HBV RNA (B) and encapsidated HBV RNA (C) were quantified by real time RT-PCR. (B) shows relative value of HBV RNA normalized by GAPDH mRNA. (D) HepG2.2.15.7 cells treated with or without 0.1% DMSO, 1 μM Bay41-4109 or 1, 5 and 10 μM nocodazole for 6 days were harvested and then pulled down with anti-polymerase antibody to detect HBV polymerase (upper) and core (lower). (E) HepG2.2.15.7 cells were treated with 10 μM nocodazole or 1 μM Bay41-4109 for 6 days or left untreated (control). Capsid and nucleocapsid-associated HBV DNA in these cells were detected with a native agarose gel electrophoresis followed by immunoblot and Southern blot, respectively. Total HBV core and actin proteins were also determined by immunoblot. (F) Capsid formation (upper) as well as total core (middle) and actin protein as an internal control (lower) were detected in HBV-infected HepG2-hNTCP-C4 cells upon treatment with or without 10 μM nocodazole or 1 μM Bay41-4109. Capsid formation was impaired upon nocodazole treatment.
