Fig. 4

Knockdown of VP16 restores the immediate early innate antiviral signaling triggered by MAVS-Pex or HSV-1 infection. a HEK293T cells were transfected with VP16-Flag, shNC or shRNA-VP16 as indicated for 36 h and subjected to WB analysis with anti-Flag antibody. b WT HEK293, shNC and shVP16-expressing HEK293 cells were subjected to WB analysis with anti-viperin antibody. (c, d and e) ShNC and shVP16-expressing HEK293 cells were pretreated with 50 ng/ml BFA and transfected with the indicated plasmids for 18 h in the presence of BFA, and subjected to RT-PCR (c), WB (d) and qRT-PCR (e) analysis. (f, g and h) HEK293 cells were pretreated with 50 ng/ml BFA and infected with or without WT HSV-1 at an MOI of 0.5 (+) or 1 (++). At 2 h post infection, cells were transfected with shNC or shVP16 plasmids as indicated in the presence of BFA. At 18 h post transfection, cells were harvested and subjected to RT-PCR (f), WB (g) and qRT-PCR (h) analysis. The data represent results from one of the triplicate experiments. Statistical analysis was performed using Student’s T-test with the GraphPad Prism 5.0 software. (* 0.01 < P<0.05; ** 0.001 < P<0.01; *** 0.0001 < P<0.001)