Figure 4
The effect of HBeAg on regulation of SOCS2 expression and ERK signaling. (A) HepG2 cells were transfected with pCMV-HBeAg at different concentrations for 24 h. SOCS1, SOCS2, and SOCS3 mRNAs were measured by real-time PCR (upper). SOCS2 and β-actin proteins were detected by Western blot analyses (lower). (B–E) HepG2 cells were incubated with rHBeAg at different concentrations (B) or transfected with pCMV-HBeAg for different times (C). HepG2 cells were transfected with pCMV-Tag2B, pCMV-HBeAg, or pCMV-HBeAg-1896mut for 24 h (D) or incubated with PBS, rHBeAg, heat-inactivated HBeAg, or rHBcAg for 12 h (E). SOCS2 mRNAs were measured by real-time PCR (upper). SOCS2 and β-actin proteins were detected by Western blot analyses (lower). (F) HepG2, pBlue-SK-transfected HepG2, pHBV1.3-transfected HepG2, or HepG2.2.15 cells were harvested 24 h after transfection. SOCS2 mRNAs were measured by real-time PCR (upper). SOCS2 and β-actin proteins were detected by Western blot analyses (lower). (G) HepG2 cells were transfected with pBlue-SK or pHBV1.3 for different times. SOCS2 mRNA was measured by real-time PCR (upper). SOCS2 and β-actin proteins were detected by Western blot analyses (lower). (H) HepG2 cells were transfected with pCMV-tag2B or plasmids expressing individual HBV proteins for 48 h. SOCS2 mRNA was determined by real-time PCR (upper). SOCS2 and β-actin mRNAs were determined by semi-quantitative RT-PCR (lower). (I) HepG2 cells were transfected with pBlue-SK, pHBV1.3, or pHBV1.3-1896mut for 24 h. SOCS2 mRNA was determined by real-time PCR (upper). SOCS2 and β-actin proteins were detected by Western blot analyses (lower). (J) HepG2 cells were pretreated with each of specific kinase inhibitors for 12 h, and incubated with rHBeAg or PBS for 24 h. SOCS2 mRNAs were measured by real-time PCR. (K) HepG2 cells were incubated with PBS, rHBeAg, or 12-O-tetradecanoylphorbol 13-acetate (TPA) for 12 h. p-ERK, ERK, and GAPDH proteins were detected by Western blot analyses. Data shown are means ± SE; n = 3. *p < 0.05.
