K-63 polyubiquitination of C417R mutated NEMO is impaired in EDI MoDCs stimulated through CD40 but not through TLR4. (A) Polyubiquination of NEMO in response to CD40L (2.5 μg/mL) for the indicated time periods in normal and EDI MoDCs. (B) Polyubiquitination of NEMO when stimulated with LPS (2.5 μg/mL) for the indicated time periods in normal and EDI MoDCs. Cell lysates were immunoprecipitated (IP) with anti-NEMO monoclonal antibody and then analyzed with an anti-Ub antibody by immunoblotting (IB). The blot was reprobed with an anti-IKKβ antibody that served as a loading control.