Skip to main content
. 2015 Feb 5;96(2):266–274. doi: 10.1016/j.ajhg.2014.11.019

Figure 3.

Figure 3

DDX58 Mutations Constitutively Activate RLR-IFN Production Pathways

(A) In HEK293FT cells, FLAG-DDX58 (WT), FLAG-DDX58-p.Glu373Ala, or FLAG-DDX58-p.Cys268Phe was transfected along with reporter construct pLuc-PRDIII-I or pLuc-NF-κB for 24 hr. A dual-luciferase assay was performed after 12 hr of polyI:C (1 μg/ml) stimulation.

(B and C) HEK293FT cells were transfected with the indicated plasmids for 30 hr and then stimulated with polyI:C (1 μg/ml) for the indicated time. After stimulation, the lysates were separated by SDS-PAGE (B) or on native gels (C). Immunoblot assays were performed with the indicated antibodies.

(D) HEK293FT cells were transfected with the indicated plasmids for 24 hr. After stimulation with polyI:C (1 μg/ml) for 12 hr, the expression of IFNB1, TNF, ISG15, and CCL5 was measured by quantitative real-time PCR and normalized to β-actin expression. The primers, which were used for quantitative real-time PCR, were described previously.13 Statistical analysis using t tests was conducted in GraphPad Prism 4 (∗p < 0.05, ∗∗p < 0.005, ∗∗∗p < 0.001).