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. 2015 Oct 12;125(11):4239–4254. doi: 10.1172/JCI81203

Figure 5. STK4 binds to and phosphorylates IRAK1.

Figure 5

(A) HEK293T cells overexpressing HA-GFP, HA-IRAK1, Myc-STK4, or Myc-K59R were prepared for immunoprecipitation and immunoblotting with the indicated antibodies. (B) Immunoprecipitation was performed using anti-IRAK1 antibody from RAW264.7 cells, followed by immunoblotting with anti-IRAK1 or anti-STK4 antibodies. (C) HEK293T cells were transfected with HA-IRAK1 and Myc-STK4 or the C-terminal deletion mutant Myc-STK4-ΔC, followed by immunoprecipitation and immunoblotting with the indicated antibodies. (D) HEK293T cells were transfected with Myc-STK4 and HA-IRAK1 or its truncated mutants, including the N-terminal fragment (100-750aa), the kinase domain (199-562aa), and the C-terminal end (562-750aa). Immunoprecipitation using anti-Myc antibody was performed followed by immumoblotting. (E) Immunoprecipitation using anti-IRAK1 antibody was performed from WT and Stk4–/– BMMs, followed by anti-IRAK1 immunoblotting. Samples were run on the same gel but not contiguous, as indicated by the black line. (A–E). Data represent the representative experiments from at least 3 independent experiments.