FIG 6.

Exogenous addition of NS3 completely restored viral growth in the presence of viperin. (A) HEK 293 cells were transiently transfected with plasmids expressing viperin and TBEV Hypr NS3, as indicated, and infected with Hypr (MOI, 0.1) for 24 h. Titers were analyzed using a focus-forming assay. (B) 293T FLP-IN T-Rex cells inducibly expressing viperin were induced with tetracycline as indicated, transfected with a plasmid expressing ZIKV NS3, and infected with ZIKV MR766 (MOI, 0.1). Supernatants were harvested 24 h p.i., and virus titers were determined using a focus-forming assay. (C and D) HEK 293 cells were transiently transfected with plasmids expressing viperin and JEV NS3 (C) or ZIKV MR766 NS3 (D), as indicated, and were infected with Hypr (MOI, 0.1) for 24 h. Titers were analyzed using a focus-forming assay. The total DNA amount was kept constant with empty vector. Means ± SEM of data from three independent experiments are shown. *, P < 0.05; **, P < 0.01; ***, P < 0.001; ****, P < 0.0001 (calculated with unpaired Student t test).