Figure 5.

The five host factors affect the abundance of multiple HBV molecules

(A) Kinetics of the decrease in extracellular HBsAg produced by HepG2.2.15 cells transfected with siRNAs. The amount of HBsAg produced was measured in 12-h intervals after siRNA transfection and normalized to the level produced by mock-treated cells. The siRNAs were transfected at a final concentration of 10 nM. REP 2139 was at 20 nM. (B) siRNAs that knock down the five host factors reduced the levels of intracellular HBsAg. The western blot image contains the signal for the large S antigen. The LC is of the β-actin protein. The knockdown of the host factors is shown in Figure S3. (C) The five host factor siRNAs can reduce the HBV polymerase levels. The western blot image shows the amounts of the HBV polymerase identified by a monoclonal antibody specific to the HBV polymerase. The L.C. shows the amount of β-actin protein. Additional western blots of cellular proteins are shown in Figure S6. HepG2.2.15 cells were transfected with a final concentration of10 nM siRNA, 5 nM ALG-20002, 8 nM ALG-10000, or 20 nM REP 2139. (D) The host factors have differential effects on the amounts of HBsAg, HBeAg, and core. ALG-10000 was transfected into HepG2.2.15 cells at 8 nM and the siRNAs at 10 nM. (E) Total HBV RNAs are modestly reduced by the knockdown of SRSF1 and HNRNPA2B1, but not by knockdown of RPLP1 or RPLP2. The cells were harvested 76 h after transfection. The amount of HBV RNA was normalized to the amount of GAPDH in the cells. In the graphs on panel A and D, each error bar represents one standard deviation of uncertainty.