FIG 7.

Silencing of RXRα and PLA2G2A modestly increases the level of cccDNA. (A) HBV cccDNA was isolated by the Hirt method and confirmed by Southern blot analysis. Lane M, molecular mass marker. (B) Knocking down RXRα and PLA2G2A expression in HepG2-NTCP (AC12) cells prior to viral inoculation enhanced HBV RNA and cccDNA formation. Cells were transfected with siRNAs against RXRα, PLA2G2A, or a control siRNA. At 3 days posttransfection, cells were incubated with HBV for 24 h; total RNA and cccDNA were extracted from infected cells at 7 dpi and detected by Northern and Southern blot analysis, respectively. (C) HBV cccDNA was detected in infected HepG2-NTCP (AC12) cells treated with bexarotene (0.5 or 0.25 μM) or DMSO during viral inoculation. (D) Total DNAs were isolated from infected cells at the indicated times, and HBV cccDNA were quantified by real-time qPCR assays. (E) AA treatment during HBV incubation reduced HBV cccDNA formation in HepG2-NTCP cells. Cells were treated with different doses of AA (or methanol control [Me]) during HBV incubation for 24 h (coincubation) or treated with AA after HBV inoculation (postinoculation). Total RNA or HBV cccDNA was extracted and detected by Northern and Southern blot analysis.