FIG. 2.

Inhibitory effects of engineered miR122 target sites on E1A protein (A) and mRNA (B) expression in infected liver cells. (A) Huh7 and A549 cells were infected with Ad5/3-Δ24 (Ad5/3) or Ad5/3-122 (122) at a MOI of 0.05 or left uninfected (−). Three days later, the cells were harvested and analyzed by Western blotting for E1A protein expression (top panels) and total immunoreactive adenoviral protein expression (bottom panels). α-E1A, anti-E1A antibody; α-Ad, antiadenovirus antibody. (B) Real-time RT-PCR was used to quantify E1A mRNA from total RNA extracted from Huh7 cells infected with Ad5/3-Δ24 or Ad5/3-122 at a MOI of 0.05. The relative expression of E1A mRNA at 1, 2, and 3 days (d) postinfection with these two viruses was determined based on an E1A mRNA dilution standard and the parallel quantification of GAPDH mRNA from the same specimens.