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. 2014 May 25;2014:420658. doi: 10.1155/2014/420658

Figure 5.

Figure 5

EAV nsp1 inhibits the NF-κB signaling pathway. Suppression of (a) TNF-α, (b) MAVS, (c) TRIF, (d) IKKβ, and (e) p65 subunit of NF-κB-induced NF-κB-dependent reporter gene expression by EAV nsp1 in HEK293T cells. (a) Cells cultured in 24-well plates were cotransfected with pNF-κB-Luc, pRL-SV40, and pCAGGS expressing nsp1 or pCAGGS empty vector for 24 h and subsequently stimulated with TNF-α (20 ng/mL) for 6 h. (b)–(e) Cells were cotransfected with plasmid pEGFP-N1-MAVS, pcDNA3-TRIF, pCMV2-IKK2-WT, or pEGFP-N1-p65 along with a pCAGGS expressing nsp1 protein or pCAGGS empty vector, pRL-SV40, and a luciferase reporter plasmid pNF-κB-Luc for 24 h. Cells were harvested and measured for firefly and Renilla luciferase activities. Relative luciferase (RLU) activity is defined as a ratio of firefly luciferase reporter activity to Renilla luciferase activity. Mean value ± SEM from three experiments is shown.