Figure 2

SMARCA2 is required for ISG induction but not for expression of MxA. (a) A549-wt cells were transfected with a non-targeting (NT) siRNA, or siRNAs targeting MxA (MX1), JAK1, or SMARCA2 (GE Dharmacon D-017253-01). 48 h post transfection the cells were either treated, or not, with IFN-α (1000 U/mL) and 24 h later were infected with H7N7-RL at an MOI of 0.3. Luciferase activity was measured 24 h post infection to determine virus reporter activity or cells were lysed and subjected to western blot analysis of the indicated proteins. Full-length blots are presented in Supplementary Figure S8c. Virus reporter activity was determined using 6 technical replicates. All data were normalized to the respective NT control. Error bars indicate the standard error of the mean of three independent experiments. Student’s t-test was performed to determine the P value. *P < 0.05, **P < 0.01. (b) A549-wt cells were treated as described in (a) but infected at an MOI of 1 for 24 hours. For immunofluorescence analysis (upper panel) MxA (red) and NP (green) were stained with specific antibodies. DAPI was used to counterstain the nucleus (blue). Lower panel: The number of cells positive for NP and MxA was quantified using flow cytometry analysis.