Fig 2.

Analysis of the F proteins of the N-glycosylation mutants by Western blotting and PNGaseF digestion. Vero cells were infected with wild-type rBC and N-glycosylation mutant viruses at an MOI of 1, and total proteins were collected after 36 h p.i. Samples were resolved on 10% polyacrylamide gels in the presence of reducing agent. Samples from rBC, rNg1, rNg2, and rNg3 (A) and rBC, rNg4, rNg5, and rNg2 + 5 (B) were digested overnight with PNGaseF or kept as untreated controls, separated by 10% polyacrylamide gels in the presence of reducing agent, and blotted onto nitrocellulose membranes. Western blot analysis was performed using a rabbit antiserum raised against a synthetic peptide designed from the F protein cytoplasmic tail. D, digested; UD, undigested.