FIG. 6.

A3G, A3F, and A3B are not required for the antiviral effect of IFNs against HBV. Doubly transduced HepG2-H1.3 cells were treated every 2 days with IFN-γ plus TNF-α, IFN-α plus TNF-α, or the RT inhibitor 3TC as indicated and collected after 6 days for total RNA, protein, and core-associated HBV DNA analyses. ND, not determined. (A) Levels of A3G, A3F, and A3B gene expression were assessed by quantitative real-time RT-PCR. Quantities were normalized using TFRC and TBP and expressed as change (n-fold) relative to the baseline levels obtained from untreated control HepG2-H1.3 cells (Vif _C133S shCtrl). Values represent means ± standard deviations for independent duplicates. (B) Aliquots of the lysates were subjected to Western blot analysis using anti-A3G, anti-Vif, and anti-PCNA antibodies. Each lane is a pool of independent duplicates. Molecular masses in kDa are given to the right. (C) Core-associated HBV DNA was purified from the remaining lysate and analyzed by quantitative real-time PCR. Levels are expressed relative to the amounts of core-associated HBV DNA produced in the different untreated HepG2-H1.3 cell lines, which were given the arbitrary value of 100%. Values represent means ± standard deviations for independent duplicates. Data are from one independent experiment representative of two. WT, wild type.