FIG. 3.

Reactivities of polyclonal antibodies to GP and SGP in Western blot and RIP assays. (A) Paired lanes of untreated (−) and endoglycosidase F/N-glycosidase F-digested (+) virion proteins labeled with [³⁵S]cysteine, run on 10% gels, and blotted onto nitrocellulose membranes. Blots were directly exposed to X-ray film (³⁵S) to identify locations of structural proteins and reacted in enzyme immunoassays (chemiluminescent) with guinea pig antisera. Arrows indicate the decreased size of virion GP1 and GP2 glycoproteins due to removal of N-linked glycans. Sera were derived from animals immunized with naked plasmid DNA which directed the expression of GP (α-GP), SGP (α-SGP), or vector-only (α-VEC) products. Asterisks identify SGP and deglycosylated SGP detected by α-SGP. Shown in panel B are lanes containing [³⁵S]cysteine-labeled proteins from an EBO virion preparation (marker) and RIP products immunoprecipitated from supernatant fluids depleted of virions (pelleted through 20% sucrose cushion) with the same antisera as used for panel A.