FIG. 4.
Coprecipitation of TCRζ with nef from Cos 18 cells. Cos 18 cells expressing CD8-TCRζ fusion protein were mock transfected (mock) or transfected with plasmids encoding full-length (SIVmac239nef) or truncated (Δnef1 to Δnef3-4) SIVmac239 nef. nef immune complexes were precipitated with anti-AU1 monoclonal antibody, washed extensively, resolved by SDS-12% PAGE, and transferred onto a membrane filter. CD8-TCRζ coimmunoprecipitated with nef was detected by the anti-TCRζ monoclonal antibody (upper panel). Expression of the full-length or the truncated SIVmac239 nef proteins in the corresponding transfected Cos 18 cells was detected by separating the whole-cell lysates in an SDS-12% polyacrylamide gel, transferring onto a nylon membrane, and probing with the anti-AU1 monoclonal antibody (lower panel).