Figure 2
The role of HBeAg in regulation of IFN receptors expression, TYK2 stability and TYK2 phosphorylation induced by IFN-α and IFN-λ1. (A) HepG2 cells were transfected with pCMV-Tag2B or pCMV-HBeAg for 48 h. Cells were Fc-blocked by treatment with human IgG prior to staining. 5 × 105 to 1 × 106 cells in PBS buffer were incubated with specific antibody and analyzed by flow cytometry using a FACSCalibur (Beckman Coulter) to detect the levels of IFN-α/βR1 (a), IFN-α/βR2 (b), IL-28R1 (c), and IL-10Rβ (d) proteins. (B) HepG2 cells were incubated with PBS or rHBeAg at 50 ng/ml for 24 h, and treated with rhIFN-α at 300 U/ml or rhIFN-λ1 at 100 ng/ml for 30 min. Cells were harvested and lysed, and p-TYK2, TYK2, p-JAK1, JAK1, and GAPDH proteins in the cell lysates were detected by Western blot analyses. (C) HepG2 cells were transfected with pCMV-Tag2B or pCMV-HBeAg for 12 h, and incubated with anti-HBeAg (10 μg/ml) for another 36 h. Cells were treated with rhIFN-α at 300 U/ml or rhIFN-λ1 at 100 ng/ml for 30 min before harvest. Cells were lysed, and p-TYK2, TYK2, p-JAK1, JAK1, and GAPDH proteins in the cell lysates were detected by Western blot analyses.
