Inhibition of hepatitis B virus surface antigen expression by small hairpin RNA in vitro

Figure 1 Structures of hairpins and U6 expression cassette. A and B: Short-hairpin structures of pAVU6+4sh421 and pAVU6+4sh81, respectively. The loop was designed with TTCG. C: Map of pAVU6+27 U6 expression cassette including U6 promoter, the first 27 nucleotides of human U6 RNA, inserted fragments and polymerase III stem terminator.

Figure 2 Electrophoresis analysis of digestion using BamH I. Lanes 1 and 2: pHBS-GFP plasmid DNA; lane 3: DNA marker pHBS-GFP.

Figure 3 Electrophoresis analysis of PCR products. Lane 1: pAVU6-4sh421 vector; lane 2: pAVU6-4sh81 vector; lane 3: pAVU6+27 vector; lane 4: DNA marker.

Figure 4 Flow cytometric detection of HBS-GFP fusion protein in HepG2 cells. HepG2 cells were transfected with pAVU6+4sh421 and pAVU6+4sh81 vectors and analyzed by flow cytometry at 72 h post-cotransfection. The values obtained from three independent experiments were expressed as mean±SD.

Figure 5 HBV cDNA copies detected by real time PCR analysis. Data showed shRNA-mediated inhibition of expression of HBs-GFP gene. Positive, mock, and negative groups represented as pHBS-GFP DNA, cDNA of cells only transfected with pHBS-GFP, respectively.

Figure 6 HBsAg concentration in the supernatant of shRNA-treated HepG2. 2.15 cells.

Figure 7 HBeAg concentration in the supernatant of shRNA-treated HepG2. 2.15 cells (S/N ratio, signal-to-noise ratio).