FIG. 2.
Presence of the minor envelope proteins in viral particles. BHK-21 cells were transfected with in vitro transcripts derived from the EAV cDNA clones WT, KO-GP2b, KO-GP3, or KO-GP4. At 7 h p.t., the cells were labeled with [35S]cysteine for 12 h. After removal of cell debris by low-speed centrifugation, the viral particles present in the culture medium were dissolved in lysis buffer and subjected to immunoprecipitation with αE, αGP2b, αGP3, or αGP4 in the presence of 5 mM dithiothreitol. The immunoprecipitates were analyzed under reducing conditions by SDS-20% PAGE (αE) or SDS-15% PAGE (αGP2b, αGP3, and αGP4). The values on the left are the molecular sizes, in kilodaltons, of marker proteins analyzed in the same gel.