Figure 6. ZAP interacts with HBV RNA through its zinc finger motifs.

HepG2 cells in 35 mm dishes were cotransfected with 3 µg of pCMVHBV, and 1 µg of control vector, or ZAP-S, or ZAP-SΔ4ZFs. Cells were harvested at day 3 post transfection. One set of cell samples was used to analyze HBV RNA and ZAP proteins as input controls. Another set of cells were lysed with cell lysis buffer containing RNase inhibitors. Immunoprecipitations were performed with beads covalently coated with anti-HA antibodies. Bonded HBV RNA and ZAP proteins were analyzed by Northern blot and Western blot assays, respectively (see Materials and Methods for details). Beads coated with anti-FLAG antibodies served as negative control (data not shown).