Extended Data Fig. 6: Role of TRAFs and PKC in TBK1 phosphorylation and activation.
From: TBKBP1 and TBK1 form a growth factor signalling axis mediating immunosuppression and tumourigenesis
a,b, Immunoblot analysis of the indicated phosphorylated (p-) and total proteins in whole-cell lysates of EGF-stimulated A549 cells stably infected with a control shRNA (shCtrl) or two different shRNAs targeting Traf6 (a) or Traf3 (b). c,d, Immunoblot analysis of the indicated phosphorylated (p-) and total proteins in whole-cell lysates of A549 cells stimulated with EGF in the presence of the PKC inhibitor GF109203X or solvent control DMSO. e, Immunoblot analysis of the indicated phosphorylated (p-) and total proteins in whole-cell lysates of control or PKCe-knockdown A549 cells, stimulated with EGF for the indicated time periods. f,g, Immunoblot analysis of T538-phosphorylated (p-) and total PKCq in the lysates of A549 cells stimulated with insulin (f) or FBS (g). h,i, immunoblot analysis of TBK1 and S172- or S716-phosphorylated (p-) TBK1 following HA IP from A549 cells stably transduced with HA-TBK1 and stimulated with either EGF (h) or Insulin (i). j, Immunoblot analysis of S172-phosphorylated (p-) and total TBK1 in whole-cell lysates of HEK293T cells transfected with the indicated expression vectors. k, CoIP assays to detect TBK1 dimerization based interaction of HA-TBK1 with Flag-TBK1 (upper) and immunoblot analysis of protein expression level in lysates (lower). l, Immunoblot analysis of the indicated phosphorylated (p-) and total proteins in LPS-stimulated control shRNA-transduced A549 cells or Tbk1 shRNA (D5)-transduced A549 cells that were reconstituted with and an empty vector or shRNA-resistant expression vectors encoding HA-tagged TBK1 wildtype (WT) or S716A. Data are representative of three independent experiments. Unprocessed blots are shown in Unprocessed Blots Fig. 6.
