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. 2019 Feb 26;10:290. doi: 10.3389/fimmu.2019.00290

Figure 4.

Figure 4

HSV-2 ICP27 inhibits IFN-β production through IRF3 signaling pathway. (A) HSV-2 ICP27 inhibits the IRF3 promoter element PRD(III-I)4. HEK 293T cells were seeded in 24-well plates and co-transfected with empty vector pcDNA3.1(+), plasmid expressing HSV-2 ICP27 or influenza virus NS1, together with PRD(III-I)4-Luc and the internal control phRL-TK. At 24 h post-transfection, cells were stimulated with or without 100 HAU ml−1 SeV for 16 h and lysed for DLR assay. (B–F) HSV-2 ICP27 inhibits RIG-I, MAVS, TBK1, IKK-ε, or IRF3-5D induced IFN-β promoter activation in a dose-dependent manner. HEK 293T cells were seeded in 24-well plates and co-transfected with empty vector pcDNA3.1(+) or HSV-2 ICP27 expressing plasmid, and plasmid expressing IRF3 signaling pathway component RIG-IN, MAVS, TBK1, IKK-ε, or IRF3-5D, together with the reporter plasmid p125-Luc and the internal control phRL-TK. At 40 h post-transfection, cells were lysed for DLR assay. At the same time, cells were lysed for Western Blot. RIG-I, MAVS, TBK1, IKK-ε, and IRF3-5D were detected by Anti-FLAG Ab while actin was detected by Anti-actin Ab. The data shown are representative of three independent experiments, with each condition performed in triplicate (mean ± SD) (A–F). *P < 0.05; **P < 0.01; ***P < 0.001; ****P < 0.0001. For Western Blot, one representative experiment out of three is shown (B–F).