Figure 5. p62 inhibits the interaction of TRAF6-ECSIT and inhibits the ubiquitination of ECSIT. (A) HEK293T cells were transfected with mock, Flag-TRAF6, Myc-ECSIT, and Myc-p62, as indicated. At 38 h post transfection, the transfected cells were extracted and cell lysates were subjected to immunoprecipitation with anti-Flag antibody followed by IB using anti-Flag, anti-p62, or anti-ECSIT antibody. (B) HEK293T cells were transfected with mock, Flag-TRAF6, Myc-ECSIT, and different concentrations of Myc-p62, as indicated. At 38 h post transfection, transfected cells were extracted and cell lysates were subjected to immunoprecipitation with anti-Flag antibody followed by IB using anti-Flag, anti-p62, or anti-ECSIT antibody. (C) HEK293T cells were transfected with mock, Flag-TRAF6, Myc-ECSIT, HA-Ub, and different concentrations of Flag-p62, as indicated. At 38 h post transfection, transfected cells were extracted and cell lysates were subjected to immunoprecipitation with anti-Myc antibody followed by IB using anti-HA, anti-Myc, anti-p62, or anti-TRAF6 antibodies.