Sustained interaction of TANK with a nondegradable TRAF3 deletion mutant but not endogenous TRAF3 following CD40 ligation.

KMH2 cells and δ300TRAF3 transfectants were metabolically radiolabeled in culture and stimulated with CD40L for the indicated times prior to lysis. TRAF3 was immunoprecipitated from untransfected and transfected KMH2 cell extracts using antihuman TRAF3 antibody and anti-FLAG antibody, respectively. This was followed by TANK immunoprecipitation from either the TRAF3 immunoprecipitates (A) or FLAG immunoprecipitates (B), using anti-human TANK antibody. The position of TANK is indicated by the arrow.