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. 2016 Dec 12;198(2):808–819. doi: 10.4049/jimmunol.1601009

FIGURE 4.

FIGURE 4.

FBXO17 interacts with IRF3 specifically during IFN-I signaling pathway. (A) The luciferase activity of HEK293T cells transfected with the FBXO17 expression plasmid, an ISRE luciferase reporter, and internal control Renilla luciferase reporter, plus RIG-I (2 CRAD), MAVS, TBK1, IKKi, or IRF3. (B and C) HEK293T cells were transfected with the indicated expression plasmids. The cell lysates were immunoprecipitated with anti-HA or anti-FLAG, and immunoprecipitates were analyzed by immunoblotting with the indicated Abs. (D and E) The cell lysates prepared from A549 cells were immunoprecipitated with anti-IRF3 or anti-FBXO17. The immunoprecipitates were analyzed by immunoblotting with the indicated Abs. (F) HA-IRF3 was cotransfected with FLAG-FBXO17 truncated mutants into HEK293T cells. The Co-IP was performed and analyzed by immunoblotting with anti-HA Ab. (G) The luciferase activity of HEK293T cells transfected with FBXO17 truncated mutants, an IFN-β luciferase reporter and internal control Renilla luciferase reporter, followed by SeV infection for 12 h. (H) HA-FBXO17 was transfected with FLAG-IRF3 truncated mutants into HEK293T cells. The Co-IP was performed and analyzed by immunoblotting with anti-HA Ab. (I) HA-FBXO17 FBA domain was transfected with IAD truncated mutants of IRF3 with a FLAG tag into HEK293T cells. The Co-IP was performed and analyzed by immunoblotting with anti-HA Ab. All results are representative of three replicate experiments. Student t test. ***p < 0.001, ****p < 0.0001. DBD, DNA-binding domain; F-BOX, F-box domain.