FIG 3.

DMXAA-induced antiviral response in macrophages reduced the HBV nucelocapsids in a hepatocyte cell line. AML12HBV10 cells cultured in the absence of tetracycline for 1 day were either directly treated with the indicated concentrations of DMXAA, poly(I:C), or Pam3CSK4 or indirectly treated with 50% of the conditioned media harvested from RAW264.7 cells (treated with the indicated concentrations of DMXAA or TLR agonists for 12 h). The AML12HBV10 cells were harvested 2 days after treatment for the following analyses. (A) Intracellular HBV RNA was determined by Northern blotting. pgRNA, pregenomic RNA; envRNAs, viral mRNAs specifying envelope proteins. 18S rRNA served as a loading control. (B) HBV core protein was determined by Western blot assay with total cell lysate. β-Actin served as a loading control. The amounts of HBV capsids (C) and capsid-associated HBV DNA (D) were determined by a native agarose gel assay. (E) Cytoplasmic HBV core DNA was determined by Southern blot hybridization.