Fig. 5

NS1 variants differentially modulate type I IFN response. CLEC213 cells in 24-well plates were transfected with the two plasmids used for the dual-luciferase assay for IFN-induced chicken Mx promoter activity, along with recombinant expression vectors encoding the indicated NS1-variants. Cells were stimulated at 24 h post-transfection by a L-poly(IC) treatment, and subsequently lysed 24 h later (48 h post-transfection). The chMx promoter-driven Firefly activity (FFL) was measured in the lysates (b) and normalized relative to the Tk-promoter driven Renilla-luciferase activity (a). (c) Fold-change ratios (induced/non-induced) are expressed relative to that calculated from the respective empty-vector control value set at 100%; * P < 0.05; *** P < 0.001 in a Dunnett’s test comparing each set to the wt-NS1 condition. (d) Western-blot detection of NS1 in transfected cells. Proteins in the lysates (1/8th of the 100 μl-lysate) were separated through SDS-PAGE, then transferred to a nylon membrane and revealed as in Fig. 3. Data are from five independent experiments, each point in (a) and (b) representing the geometric mean of a technical triplicate