Figure 3
From: STRAP Acts as a Scaffolding Protein in Controlling the TLR2/4 Signaling Pathway
STRAP associates with TAK1, IKKα, and p65.
(a–c) STRAP interacts with TAK1, IKKα, and p65. RAW 264.7 cells stably expressing GFP-STRAP were treated with LPS (80 ng/ml) at different time points. Cell lysates were immunoprecipitated with anti-TAK1 (a), anti-IKKα (b), or anti-p65 (c) antibodies and immunoblotted with anti-GFP antibody. (d–f) STRAP endogenously binds to TAK1, IKKα, and p65. After LPS stimulation, cell lysates were immunoprecipitated with anti-TAK1 (d), anti-IKKα (e), or anti-p65 (f) antibodies and immunoblotted with anti-STRAP antibody. (g) The interaction between IKKα and TAK1 or IKKα and p65 was increased by STRAP overexpression. The association of IKKα with TAK1 or p65 was determined by immunoprecipitation with anti-IKKα antibody and immunoblotting with the indicated antibodies. (h) The interaction of IKKα with p65 or TAK1 was reduced by STRAP depletion. Lysates from LPS-stimulated RAW macrophages stably expressing shRNA-GFP or shRNA-STRAP were immunoprecipitated with anti-IKKα antibody and immunoblotted with the indicated antibodies. (i) TAK1 endogenously interacts with p65 and its interaction is reduced by STRAP depletion. Lysates from LPS-stimulated macrophages were immunoprecipitated with anti-p65 antibody and immunoblotted with anti-TAK1 antibody. Data are representative of three independent experiments.
