Figure 4

Figure 4

Physical interaction of zbTRIM25 with zbRIG-I. (A) Schematic representation of full-length zbRIG-I and zbRIG-I deletion mutants. (B–E) Interactions of GFP-tagged zbRIG-I-2CARD (B), zbRIG-I-Δ2CARD (C), zbRIG-I-RD (D), or zbRIG-I-Δ(CARDs+RD) (E) with Flag-tagged zbTRIM25 were examined using immunoprecipitation assays. HEK 293T cells were transfected with plasmids as indicated for 24 h. After treated with poly I:C for 24 h, cells were lysed, and the cell lysates were either analyzed directly by using anti-GFP and anti-Flag antibodies via Western blotting (Input) or subjected to immunoprecipitation using anti-GFP antibodies. The precipitates (IP) were analyzed by Western blotting with anti-GFP and anti-Flag antibodies, respectively. (F) Schematic illustration of zbTRIM25 truncations. Interactions of GFP-zbRIG-I-RD with Flag-zbTRIM25-ΔSPRY (G) or Flag-zbTRIM25-SPRY (H), GFP-zbRIG-I-2CARD with Flag-zbTRIM25-ΔSPRY (I), or Flag-zbTRIM25-SPRY (J) were examined using immunoprecipitation assays. HEK 293T cells were transfected with plasmids as indicated for 24 h. Cells were lysed after treated with poly I:C for 24 h. Immunoprecipitation and immunoblotting were performed with indicated antibodies.