(a) Evaluation of Tbk1, Mavs and Tmem173 knock-out in D4C9 cells overexpressing Egfp or Fbxw7(wt) by western blot. (b-c) Relative Ifnb1 mRNA expression in D4C9 cells overexpressing Egfp or Fbxw7(wt) and transduced with a sgRNA targeting Tbk1, Mavs, Tmem173, or a non-targeting sgRNA, following transfection with (b) LMW poly(I:C) (1μg/ml) or (c) poly(dA-dT) (1μg/ml) for 24h. Each dot represents a biological replicate. One-way ANOVA with Sidak’s multiple comparisons test was used to determine statistical significance. *, P <0.05; ****, P <0.0001. (d) Evaluation of Ifih1 and Ddx58 knock-out in D4C9 cells overexpressing Egfp or Fbxw7(wt) by western blot. (e-f) Relative Ifnb1 mRNA expression in D4C9 cells overexpressing Egfp or Fbxw7(wt) and transduced with a sgRNA targeting Ifih1, Ddx58, or a non-targeting sgRNA, following transfection with (e) LMW poly(I:C) (1μg/ml) or (f) HMW poly(I:C) (1μg/ml) for 24h. Each dot represents a biological replicate. One-way ANOVA with Sidak’s multiple comparisons test was used to determine statistical significance. ****, P <0.0001. (g) Protein expression of components of the viral sensing pathways in control and Fbxw7-deficient D4C9 cells. (h) Protein expression of components of the dsRNA sensing pathway in D4C9 cells overexpressing Egfp or wild-type Fbxw7(wt).