Figure 3

PRRSV Nsp2 significantly enhances NF-κB-regulated gene expression.

(A) The wild-type Nsp2 (0.6 μg) and the empty vector were transfected into HeLa cells along with various reporter plasmids (0.2 μg) and pRL-TK (0.05 μg), respectively. Luciferase reporter assays were performed at 36 h post-transfection. The results are expressed as fold induction of IL-6, IL-8, COX-2, and RANTES promoter luciferase activity relative to the basal level. (B) HeLa cells were transfected with 1 μg of plasmid encoding Nsp2, or empty vector. After 36 h incubation, total RNA was extracted and the expression of IL-6, IL-8, COX-2, RANTES, and GAPDH genes were evaluated by real-time RT-PCR. Results are expressed as increasing mRNA levels relative to those in cells transfected with the empty vector and were normalized by using GAPDH housekeeping gene expression. All data represent the means and standard deviation of three independent experiments. **P < 0.01 as compared with vector control.