FIG. 4.
Analysis of the expression and incorporation into pseudotyped retroviruses of Ebola virus GPs with substitutions of ectodomain cysteine residues. φNX cells were transfected with plasmids encoding Ebola virus GPs. The cell lysates (L) and viral particles collected from the culture medium (M) were analyzed by SDS-PAGE (8.5% acrylamide) and immunoblotting with anti-Ebola virus SGP-GP antibody. Analysis of a cell lysate aliquot that was treated with PNGase F (+), which removes N-linked glycosylation, is also shown. The migration positions of mature GP1, GP0 (the glycosylated but uncleaved form), GPpre (the N-glycosylated but not O-glycosylated uncleaved form), and deglycosylated GP0 and GPpre are indicated. WT, wild type; Neg, ΦNX cells transfected with pcDNA3 vector.