Fig. 5.
Analysis of TRAF6 polyubiquitination. HEK293T cells were mock transfected (control) or transfected with FLAG-tagged wild type (WT) or the indicated mutated forms of TRAF6 along with a vector expressing ubiquitin. The TRAF6 proteins were immunoprecipitated 16 hours post transfection and analyzed by immunoblotting using the anti-FLAG M5 monoclonal antibody or antibodies that recognize ubiquitin or K63-linked polyubiquitin chains. (A) Representative Western blot showing poyubiquitinated (poly-Ub) and non-ubiquitinated (TRAF6) TRAF6 proteins. (B) Representative Western blot showing poyubiquitinated by K63-linked chains (K63 poly-Ub) and non-ubiquitinated (TRAF6) TRAF6 proteins.