Figure 1.

Pseudorabies virus (PRV) UL24 dampens TNF-α-mediated NF-κB activation. (A) HEK293T cells and (B) HeLa cells were co-transfected with an NF-κB-Luc reporter plasmid, the pRL-TK plasmid (transfection control), along with a UL24 plasmid or an empty vector. Luciferase activity was measured at 24 h post transfection, and fold activation was determined compared to that of the empty vector (fold activation of the empty vector was set as 1). The expression of UL24 was analyzed by WB using an anti-HA MAb. (C) HeLa cells were co-transfected with 200 ng of p65 plasmid and an empty vector or an increasing amount of the UL24 plasmid (0, 20 ng, and 200 ng); the cells were lysed at 48 h post transfection, and luciferase activity was measured. The expression of UL24 was analyzed by WB using an anti-HA Mab. (D) HeLa cells were transfected with 2 μg of UL24 plasmid or an empty vector. At 24 h post transfection, the cells were mock treated or treated with TNF-α (10 ng/μL) for 8 h. Total RNA was extracted and digested with DNase I and further subjected to reverse transcription. The cDNA was used for semi-quantitative PCR to investigate the accumulation of human IL-6 and IL-8 mRNA and UL24. The data represent results from one of three independent experiments. Error bars represent standard deviations of data from three replicates of one independent experiment. * stands for p < 0.05.