Figure 1. FAK Depletion Enhances RNA Virus Replication and Attenuates Antiviral Signaling.

(A), FAK^+/+ or FAK^-/- MEFs were infected with CVB (MOI=1), ReoV (MOI=5), SeV (10 HAU/mL), or VSV (MOI=0.1) for 16 hr. Cells were fixed and stained for markers of virus infection (CVB [VP1], ReoV [polyclonal antisera], SeV [C protein]) or assessed for GFP expression (VSV-GFP). Blue, DAPI-stained nuclei. (B), FAK^+/+ or FAK^-/- MEFs were infected with CVB (MOI=1), ReoV (MOI=5), SeV (10 HAU/mL), VSV (MOI=0.1), VV (MOI=10), or HSV-1 (MOI=5) for 16 hr. Following infection, cells were fixed and stained for markers of virus infection (CVB [VP1], ReoV [polyclonal antisera], SeV [C protein]) or assessed for GFP expression (VSV-GFP, VV-GFP, HSV-1-GFP). Level of infection was calculated based on the percent of virus-infected cells over total cells, as assessed by DAPI-staining. (C, D), Plaque assays for VSV performed using FAK^+/+ and FAK^-/- MEFs (C) or CVB in HeLa cells on lysates harvested from FAK^+/+ or FAK^-/- MEFs. Top, representative wells from the 10⁻⁶ dilution of virus are shown. Bottom, quantification of viral titers. (E, F), Luciferase assays from FAK^+/+ or FAK^-/- MEFs transfected with NF-κB (C) or IFNβ (D) promoted luciferase constructs and infected with SeV (20 HAU/mL) or VSV (MOI=0.5) or transfected with poly(I:C) (100 ng) for 16 h following transfection (24 hr). (G), FAK^+/+ or FAK^-/- MEFs were transfected with 100 ng poly(I:C). Total RNA was extracted ~ 24 hr post-transfection and analyzed by quantitative RT–PCR for IkBα (left) and IFN-β (right) mRNA. (H), Luciferase assays from FAK^-/- MEFs transfected with NF-κB or IFNβ promoted luciferase constructs and either vector or FAK. Following transfection (24 hr), cells were transfected with poly(I:C) (100 ng) for 16 hr and luciferase activity measured. Data in (B-H) are shown as mean ± standard deviation. Asterisks indicate P-values of ≤ 0.05.