Figure 5: TBK1 is required for ΔN146 induced signaling in immature DCs.

(A) Effect of TBK1 inhibitor on cell viability. Immature DCs were left untreated or treated with DMSO or TBK1 inhibitor BX795 at indicated concentrations. At 14 h after treatment, cell viability was measured by the trypan blue exclusion method. The results are mean values of duplicate samples with standard deviations. (B) Effect of TBK1 inhibitor on the activation of innate immune signaling pathways triggered by ΔN146 mutant. Immature DCs were treated and infected as B. At 3 hours post infection, cells were collected and subjected to immunoblotting using antibodies against IRF3, phosphorylated IRF3 (Ser396), IκBα, phosphorylated IκBα, and β-actin respectively. (C) Effect of TBK1 inhibitor on the expression of antiviral genes and inflammatory cytokines. Immature DCs were left untreated or treated with DMSO or TBK1 inhibitor BX795 (1 μM) for 1 hour. Subsequently cells were either mock infected or infected by ΔN146 mutant (5 PFU/cell) in the presence or absence of BX795 (1 μM). At 3 and 6 h post infection, RNAs were isolated from the cells and assayed by quantitative real-time PCR for the expression of IFN-β, RANTES and IL6. The data were normalized to the level of 18 S rRNA and expressed as relative expression with standard deviations among triplicate samples.