FIG. 2.
Composition of the 66- and 45-kDa complexes. EAV-infected BHK-21 cells were labeled with l-[35S]cysteine from 6.5 to 11 h p.i. After removal of cell debris from the culture medium by low-speed centrifugation, the extracellular virions were pelleted through a cushion of 20% (wt/wt) sucrose. The pellet was dissolved in lysis buffer, and the 66- and 45-kDa complexes were immunoprecipitated with αGP2b and separated in an SDS-15% PAA gel under nonreducing conditions. Subsequently, the 66- and 45-kDa complexes were purified from the dried gel (right panel) and analyzed by SDS-PAGE under reducing (+DTT) and nonreducing (−DTT) conditions (left panel, lanes 66 kDa and 45 kDa). For comparison, the immune complexes obtained after incubation of aliquots of the solubilized virus pellet with αGP2b were run in parallel (left panel, lanes αGP2b). The numbers between the panels are the molecular masses, in kilodaltons, of marker proteins analyzed in the same gel.