Figure 2. TRIM6 is required for efficient induction of IFNβ and ISGs.
(A-G) A549 cells were transfected with a control or TRIM6-specific siRNAs. (A-E) Forty h p.t., cells were mock-treated or infected with SeV (10 HAU/ml), IAV (PR8; 2 PFU/cell), (E) a recombinant NS1 RNA binding mutant virus (R38A/K41A), or (F-G) transfected with dsRNA poly (I:C). RNA levels were quantified by RT-qPCR. (E) At 8 h p.i. with IAV, cells were harvested for immunoblot (IB). (F) At 8 h p.t. with poly I:C, cells were harvested for RT-qPCR or (G) (4 h p.t.) EMSA analysis of ISGF3 binding to ISG promoters. (H) hDCs transduced with lentiviruses expressing TRIM6 shRNAs were infected with SeV or EMCV. Cells were harvested at 6 h p.i for RT-qPCR analysis or at 24 h p.i for virus titeration by RT-qPCR or plaque assay. Values are plotted as fold induction over mock samples. Data represent mean ± SD. (I) The effect of knockdown for all four donors combined was determined by two-way ANOVA. **p < 0.01; ***p < 0.001, N.S. (not significant). See also Figure S2