Extended Data Fig. 4: TBK1 is associated with mTORC1 via Raptor.
From: TBKBP1 and TBK1 form a growth factor signalling axis mediating immunosuppression and tumourigenesis
a, Schematic of Myc-BioID (Myc-tagged BirA R118G) and Myc-BioID-TBK1 constructs cloned into the pCLXSN(GFP) retroviral vector. b, Scatter plot showing several known TBK1-binding proteins and a novel TBK1-binding protein, Raptor (also called RPTOR) identified by BioID screening. c,d, Immunoblot analysis of Raptor and Rictor pulled down by TBK1 IP (c, upper) or mTOR and TBK1 pulled down by Rictor IP (d, upper), with protein expression being monitored by direct immunoblot assays (lower), using whole-cell lysates of HEK293 cells transfected with the indicated expression vectors. e, Co-IP analysis of endogenous TBK1-Raptor and TBK1-Rictor interactions in EGF-stimulated A549 cells (upper). IgG was used as a negative control for IP. f,g, Co-IP analysis of TBK1-mTORC1 interactions (upper) and direct immunoblot analyses of the indicated proteins (lower) in control or Tbk1-knockdown A549 cells stimulated with EGF. The Lysates were subjected to IP with antibody against TBK1 (f) or Raptor (g). Data are representative of three independent experiments. Unprocessed blots are shown in Unprocessed Blots Fig. 4.
