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Fig. 5 | Virology Journal

Fig. 5

From: Porcine transmissible gastroenteritis virus inhibits NF-κB activity via nonstructural protein 3 to evade host immune system

Fig. 5

Amino acids at positions 590–1215 in Nsp3 play a vital role in the inhibition of the NF-κB signaling pathway. IPEC-J2 and HEK-293 T cells were co-transfected with pNF-κB-Luc (0.5 μg), pRL-TK (0.025 μg), and one of the following plasmids (0.5 μg): pCMV-HA-Nsp3 (1–418 aa), pCMV-HA-Nsp3 (410–601 aa), pCMV-HA-Nsp3 (590–1215 aa), pCMV-HA-Nsp3 (1168–1510 aa), or pCMV-HA. After 24 h, the cells were treated with poly (I:C), while the cells that were transfected with pCMV-HA were treated with poly (I:C) or PBS as the positive and negative controls, respectively. At 12 h post-treatment, the cell lysates were prepared and subjected to dual luciferase assays (a), the RNA was extracted from cells and the mRNA levels of IL-1, IL-6, IL-8, TNF-α and β-actin were real-time PCR (b). β-actin was used as an internal reference gene. c HEK-293 T and IPEC-J2 cells were co-transfected with pNF-κB-Luc (0.5 μg), pRL-TK (0.025 μg), and different doses of Nsp3 (590–1215 aa) eukaryotic expression plasmids (0, 0.5, 1.0, and 1.5 μg). At 24 h post-transfection, 10 μg/mL poly (I:C) was added to the cells. The cells were harvested and analyzed for luciferase activity at 36 h post-transfection. Data are presented as mean ± SD. *P < 0.05 and **P < 0.01 were considered to be statistically significant and highly significant, respectively

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