FIG. 7.
Lipid raft association of patient-derived envelopes. 293T cells expressing NA420 LN40, NA420 B33, and chimeric NA420 LN40 (B33758-856) envelopes were lysed with cold Triton X-100, and the postnuclear supernatants were subjected to sucrose density gradient centrifugation. DRM and DSM fractions were immunoprecipitated with serum from a HIV patient as described in Materials and Methods and resolved by SDS-PAGE. Envelope and Gag proteins were detected by Western blotting using Chessie 8 and 183-H12-5C MAbs, respectively. Note that the substitution of the B33758-856 sequence in the LN40 gp41 cytoplasmic domain fully rescued envelope recruitment to DRMs.