Fig. 4
HBx Trp120 and Leu123 residues are critical for HBV production in mice and HepG2-NTCP cell. a, b Southern blot analysis of HBV DNA replication (a) and Northern blot analysis of HBV transcription (b) in HepG2 cells transfected with pHBV1.3-WT (wild type), pHBV1.3-HBx(W120A/L123A), or pHBV1.3-HBx-null, respectively. c–h Analysis of HBV viral protein expression in a HBV mouse model. Expression levels of serum HBV S-antigen (HBsAg) (c) and HBV e-antigen (HBeAg) (d) and expression levels of intrahepatic HBV core antigen (HBcAg) (f) and HBsAg (h) in liver lysates from mice 3 days post hydrodynamic injection of the pHBV1.3-WT replicon, the pHBV1.3-HBx(WL/AA) replicon, or the pHBV1.3-HBx-null replicon were analyzed by chemiluminescent enzyme immunoassay (CLEIA), respectively. The expression levels of intrahepatic HBcAg (e) and HBsAg (g) in the liver tissue sections from these mice were also examined by immunostaining. Scale bar is 100 μm. The red arrows indicate the hepatic cells with HBcAg or HBsAg expression. pcDNA3.1-EGFP was co-injected with the HBV replicon as an injection marker to normalize the transfection efficiency (n = 6 biologically independent animals). i Kinetics of HBeAg and j HBsAg levels post HBV-WT (blue line), HBV-WL/AA (red line), and HBV-Xnull (green line) infection in HepG2-NTCP cell line, supernatant of untreated HepG2-NTCP cell was used as a negative control (purple line) (n = 3 biologically independent samples). The data represent mean ± SD. Significant differences between groups are indicated on the top. **P < 0.01, ***P < 0.001, ****P < 0.0001; two-tailed unpaired t -tests. Source data are provided as a Source Data file
