Fig 2: The dsDNA/CDN-binding surface of DDX41 is critical for the innate-immune responses.(A,B) Quantitative PCR analyses of IFNB1 (A) and IFIT2 (B) transcription in THP-1 cells. The cells were treated with shRNA targeting the mRNA encoding DDX41, and then transfected with vectors expressing FL-DDX41 WT or mutants. Gels were run under the same experimental conditions and are shown as cropped blots. Full-length blots with the cropping lines indicated are shown in Figure S7.

Fig 3: The DDX41 DEAD domain binds to dsDNA and CDN.(A) FL-DDX41 and its truncated variants. DDX41 contains the DEAD domain (sky blue), the HELICc domain (orange) and the Zinc finger (ZF, gray). (B) Pull-down assays of DDX41, using bio-ISD (left panel) and bio-cGG (right panel). In all of the pull-down assays, the final concentration of bio-ISD, bio-cGG, bio-ATP or bio-AMPPNP is 1 µM, unless otherwise stated. Gels were run under the same experimental conditions and are shown as cropped gels. Full-length gels with indicated cropping lines are shown in Figure S7.

Fig 4: Putative dsDNA and CDN binding sites of DDX41.(A) Docking model of the DDX41 DEAD domain bound to dsDNA. The dsDNA is colored brown. (B) Electrostatic surface potentials of the DDX41 closed form (contoured from -10 kT e-1 [red] to +10 kT e-1 [blue]). (C) Pull-down assays of the DEAD mutants and bio-ISD. (D) Pull-down assays of the DEAD mutants and bio-cGG. Gels were run under the same experimental conditions and are shown as cropped gels. Full-length gels with the cropping lines indicated are shown in Figure S7.