Figure 3.

Regulation of lncATV on the host innate immune response. (A) Huh7 cells were transfected with negative control siRNA (siNC) or siRNA targeting lncATV at 48 h prior to infection. NDV (MOI = 0.01) was inoculated and allowed for another 16 h infection to trigger innate immune response. Total RNA was isolated, and qRT-PCR was performed to measure the expressions of IFNβ, ISG15, MxA, PKR, IFIT1, and RSAD2. (B) Dimerization of IRF3 was measured by native-PAGE and Western blotting. (C) Immunoblot analysis of phosphorylated proteins in lncATV knockdown and control Huh7 cells infected with NDV for indicated hours. (D) IFNβ or ISRE promoter-driven reporter construct was co-transfected with either lncATV overexpressing plasmid or empty vector (EV) into HEK293T cells. Luciferase activities were determined in the absence or presence of SeV infection. (E,F) HEK293T cells were transfected with lncATV overexpressing plasmid or empty vector as controls and then challenged with SeV (MOI = 1) for the indicated time courses. Phosphorylated TBK1 (E) or STAT1 (F) was detected by Western blotting. (G) LncATV or control empty vector was overexpressed in Raw264.7 macrophage cells. The cells were infected with VSV (MOI = 0.5) for the indicated time courses and phosphorylated IRF3 and TBK1 were detected. Typical photographs of one representative experiment were shown. Data are representative of three independent experiments and plotted as the mean ± s.d. ^**P < 0.01, ^***P < 0.001 vs. the control group. NS, not significant.