FIG 3.
BMS-806 stabilization of gp120 association with Env complexes. (A) To evaluate the stability of the noncovalent association of gp120 with Env complexes, the detergent-solubilized, His6-tagged Envs were captured on Ni-NTA beads and analyzed by Western blotting with rabbit anti-gp120 antibody or the 4E10 anti-gp41 antibody. (B) VLPs were prepared from the supernatants of A549-Gag/Env cells expressing Gag-mCherry and HIV-1AD8 Env. Triton X-100 lysates of VLPs were incubated with DMSO or the indicated molecules. Env was then captured on Ni-NTA beads and analyzed by Western blotting, as described in the legend to panel A. (C) Triton X-100 lysates of A549 or HOS cells expressing Envs from different HIV-1 strains were incubated with Ni-NTA beads in the presence of the indicated molecules, and then the captured molecules were analyzed by Western blotting as described in the legend to panel A. Lanes M, molecular mass markers. The numbers to the left are molecular masses (in kilodaltons). The experiments for which the results are shown in panels B and C were performed at least twice, and the results of a representative experiment are shown.