Figure 1.

Schematic experimental strategy used in this study. Briefly, one microgram of pHY92-1.1x-HBV-small S (-) having a full-genome of hepatitis B virus (HBV) with a knock-out small surface open reading frame (ORF) was co-transfected with three micrograms of pIRES2-EGFP-HBV-small S expressing the sub-genome of small surface region, which has ten variants including mock and wild-type into HuH-7 cell line transiently. After co-transfection, HuH-7 cells were incubated for 2 d. Supernatant and lysed pellet were collected and used for various assays. The tests were performed in triplicate. HBsAg: Hepatitis B surface antigen; ELISA: Enzyme linked immunosorbent assay; Q-PCR: Quantitative polymerase chain reaction; ROS: Reactive oxygen species; Neo/Kan R: Neomycin and Kanamycin resistance.