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Nef can interact with the Ii chain in a dileucine-dependent manner. Immunoprecipitations were performed with HeLa or HeLa-CIITA cells transfected with Nef-STOP, Nef NL4-3, or the Nef LL164AA construct by using the Ii chain-specific MAb PIN1 covalently linked to protein A beads. Immunoprecipitates (IP) were eluted at 95°C and revealed with either anti-Nef or anti-Ii MAbs and peroxidase-labeled secondary antibodies by Western blotting.
