Figure 6.
An intimate correlation between binding of c-FLIPL to MKK7 and inhibition of the MKK7–JNK pathway. (A) Schematic diagrams of deletion mutants of c-FLIPL, and the summary of their binding and inhibition experiments. (B, E) HEK293 cells were transfected with the indicated expression vectors. After immunoprecipitation (IP) with anti-Flag antibody, co-immunoprecipitated MKK7 was detected by immunoblotting with anti-HA antibody (top panels). Expression levels of the transfected proteins were analyzed as in Figure 5A. (C, D, F, G) HEK293 cells were transfected with the indicated expression vectors. After immunoprecipitation (IP) with anti-HA antibody, the phosphorylation of MKK7 (C, F) or JNK (D, G) was analyzed by immunoblotting with anti-phospho-JNK or anti-phospho-MKK7 antibodies (top panels). Expression levels of the transfected proteins were analyzed as in Figure 4A. (H) HEK293 cells were transfected with the indicated expression vectors. After immunoprecipitation (IP) with anti-Myc antibody, co-immunoprecipitated MKK7-KM was detected by immunoblotting with anti-HA antibody (top panels). Expression levels of the transfected proteins were analyzed as in Figure 5A.