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Amino acid residue 132 is critical for interaction of HIV-2 Nef with CD3ζ. 293T cells were transfected with pCGCG vectors coexpressing GFP and CD3ζ or C-terminally HA-tagged Nef variants. At 40 h posttransfection, the cells were lysed and analyzed by Western blotting or coimmunoprecipitation analysis as described in Materials and Methods.
