Skip to main content
. 2013 Jan;87(2):1221–1231. doi: 10.1128/JVI.02421-12

Fig 1.

Fig 1

Infection of HEK293T cells with IBDV strain Lx inhibits TNF-induced expression of IFN-α, IFN-β, and NF-κB. (A to D) Replication of IBDV strain Lx in HEK293T cells. HEK293T cells were mock infected (A and B) or infected with IBDV Lx at an MOI of 10 (C and D) and incubated for 3 h at 37°C. Twenty-four hours after IBDV infection, cells were subjected to IFA staining using chicken anti-IBDV antiserum followed by FITC-conjugated rabbit anti-chicken IgG antibodies and were visualized under a fluorescence microscope. (E) Kinetics of IBDV replication in HEK293T cells. HEK293T cells were infected with IBDV at an MOI of 10. At different time points (12, 24, 48, and 72 h) after IBDV infection, the viral titers in the cell cultures were determined as TCID50 using 96-well plates. (F to H) Effects of IBDV infection on TNF-induced mRNA expression of IFN-α, IFN-β, and NF-κB. HEK293T cells (2 × 105) were mock infected or infected with IBDV at an MOI of 20. Twelve hours after IBDV infection, cells were treated with TNF-α at a final concentration of 20 ng/ml. Twelve hours after TNF treatment, mRNA expression of IFN-α, IFN-β, and NF-κB was measured by qRT-PCR using specific primers. The relative level of gene expression is calculated as follows: gene expression of IBDV-infected cells or TNF-treated cells (mock- or IBDV-infected cells)/gene expression of mock-infected controls. (I to K) Effects of IBDV infection on TNF-induced activation of IFN-α, IFN-β, and NF-κB promoters. HEK293T cells (2.0 × 105) were seeded on 24-well plates and transfected with the indicated reporter gene plasmids. Six hours after transfection, cells were mock infected or infected with IBDV at an MOI of 20. Twelve hours after IBDV infection, cells were treated with TNF-α at a final concentration of 20 ng/ml. Twelve hours after TNF-α treatment, luciferase reporter gene assays were performed. The data are normalized as in panels F to H. Results are representative of three independent experiments. Data are represented as means ± SD; n = 3. ***, P < 0.001; **, P < 0.01. Ctrl, control.