FIG. 8.

Western blot analysis of Env proteins in purified RV virions. BSR cells were infected with RV constructs at an MOI of 1 for 4 days. Clarified culture supernatants were layered over 20% buffered sucrose, and virions were pelleted via ultracentrifugation. Isolated virions were resuspended in 1× Laemmli buffer with SDS (denatured). Samples run under reduced conditions had 100 mM DTT added prior to boiling and SDS-PAGE. Membrane was probed with anti-gp120 polyclonal antibody and was developed by chemiluminescence. The relative size of each band is given at right. All samples are denatured, and samples in lanes 2, 4, and 6 are also reduced. Under reduced conditions the gp140 species dissociate and the predominant Env species in both viruses is cleaved gp120 (compare lanes 4 and 6).