Figure 3
The impacts of HBeAg on ISG expression and antiviral activity induced by IFNs. (A and B) HepG2 cells were transfected with pCMV-Tag2B, pCMV-HBeAg, or pCMV-HBeAg-1896mut for 24 h and treated with rhIFN-α (A) or rhIFN-λ1 (B) for another 24 h. Total RNA was extracted from the treated cells, and mRNA levels of IFN-stimulated genes, OAS (upper panels) and PKR (lower panels), in the RNA extracts were detected by real-time PCR. (C and D) HepG2 cells were incubated with PBS, rHBeAg at 50 ng/ml, or heat-inactivated rHBeAg for 12 h and treated with rhIFN-α (C) or rhIFN-λ1 (D) for another 24 h. Total RNA was extracted from the treated cells, and mRNA levels of OAS (upper panels) and PKR (lower panels) in the RNA extracts were detected by real-time PCR. (E and F) HepG2 cells were co-transfected with pHBV1.3 and pCMV-Tag2B or pCMV-HBeAg for 24 h and treated with rhIFN-α (upper panels) or rhIFN-λ1 (lower panels) for another 24 h. Cells were harvested and lysed, and the levels of hepatitis B s antigen (HBsAg) in culture supernatants were measured by ELISA using an HBsAg diagnostic kit (E). Cells were homogenized in lyses buffer and treated with DNase. The viral cores were precipitated by adding EDTA and polyethylene glycol and concentrated by centrifugation. HBV capsid-associated DNA was quantified by real-time PCR (F). (G and H) Huh7 cells were transfected with pHBV1.3 or pHBV1.3-1896mut for 24 h, and treated with rhIFN-α (upper panels) or rhIFN-λ1 (lower panels) for another 24 h. Cells were harvested and lysed, and the levels of hepatitis B s antigen (HBsAg) in culture supernatants were measured by ELISA (G). Cells were homogenized in lyses buffer and treated with DNase. The viral cores were precipitated by adding EDTA and polyethylene glycol and concentrated by centrifugation. HBV capsid-associated DNA was quantified by real-time PCR (H). Data shown were means ± SE; n = 3. *p < 0.05.
