Fig. 5 | Cellular & Molecular Immunology

Fig. 5

From: Influenza A virus use of BinCARD1 to facilitate the binding of viral NP to importin α7 is counteracted by TBK1-p62 axis-mediated autophagy

Fig. 5

BinCARD1 facilitates the binding of NP with IMP α7. A–D Co-IP assays were performed to examine the interaction between BinCARD1 and IMP α1, α3, α5, and β1. HEK293T cells were transfected with plasmids expressing Myc-tagged BinCARD1 and Flag-tagged IMP α1 (A), α3 (B), α5 (C), or β1 (D) individually or in combination. At 36 h p.t., cell lysate proteins were immunoprecipitated with a mouse anti-Myc mAb or a mouse anti-Flag mAb and subjected to western blotting with a rabbit anti-Myc pAb or a rabbit anti-Flag pAb. E, F A co-IP assay was performed to examine the interaction between BinCARD1 and IMP α7. HEK293T cells were transfected with plasmids expressing Myc-tagged BinCARD1 and Flag-tagged IMP α7 individually or in combination. At 36 h p.t., cell lysate proteins were immunoprecipitated with a mouse anti-Flag mAb (E) or a mouse anti-Myc mAb (F) and subjected to western blotting with a rabbit anti-Myc pAb or a rabbit anti-Flag pAb. G A co-IP assay was performed to examine the interaction between BinCARD1 and endogenous IMP α7 during IAV infection. HEK293T cells were transfected with vectors to express Flag-tagged BinCARD1, and at 36 h p.t., the cells were infected with WSN (H1N1) virus (MOI = 5) for 12 h. Cell lysate proteins were immunoprecipitated with a mouse anti-Flag mAb and subjected to western blotting with a rabbit anti-IMP α7 pAb or a rabbit anti-Flag pAb. H Western blotting analysis to determine the effect of gradually increasing BinCARD1 expression on the expression of IMP α7. HEK293T cells were transfected with vectors to express Flag-tagged IMP α7 and gradually increasing levels of BinCARD1. At 36 h p.t., cell lysate proteins were western blotted with a rabbit anti-Flag pAb and a mouse anti-BinCARD1 mAb. I Co-IP assays were performed to examine the relationship among NP, IMP α7, and BinCARD1. HEK293T cells were transfected with plasmids expressing WSNNP, Flag-tagged IMP α7, and gradually increasing amounts of Myc-tagged BinCARD1 individually or in combination. At 36 h p.t., cell lysate proteins were immunoprecipitated with a mouse anti-Flag mAb and subjected to western blotting with a rabbit anti-NP pAb, a rabbit anti-Flag pAb, or a rabbit anti-Myc pAb

Back to article page