FIG. 2.

Interaction of HIV-1 EnvCD with clathrin-associated protein complexes AP-1 and AP-2 from T-lymphocyte lysates. Equal amounts of the various immobilized GST-EnvCD fusion proteins were incubated with T-cell cytosol, and bound material was separated by SDS-PAGE. (A) The effect on AP-1 binding (top) and AP-2 binding (bottom) of deletions in the cytosolic domain of Env was assessed by immunoblotting using an antibody against either one of the large chains of AP-1 (γ-adaptin) or an antibody against the large-chain β2 of AP-2. For comparison, an aliquot of the T-cell extract was loaded in the first lane. (B) As in panel A, immobilized GST-EnvCD fusion proteins were incubated with T-cell cytosol and bound material was separated by SDS-PAGE. The effect on AP-1 recruitment of mutations in the Env tail was revealed by Western blotting using an antibody against the large chain γ of AP-1.