FIG 6.
Replication and glycosylation of nsp4 mutants. (A) Schematic of nsp4 showing the location of the nsp4 mutations. (B) DBT cells were infected with the indicated viruses at an MOI of 1 PFU/cell for 24 h. At the indicated time points, supernatants were sampled, and titers were determined by plaque assay. Error bars represent the standard errors of the means of three replicates plated in duplicate. (C) DBT cells were infected with the indicated viruses at an MOI of 10 PFU/cell. Cells were starved with DMEM lacking Cys and Met in the presence of ActD for 1 h prior to being radiolabeled with [35S]Cys-Met for 2 h before lysates were harvested. Lysates were immunoprecipitated with antibodies specific to nsp4 in the presence or absence of endo-H, and proteins were resolved by SDS-PAGE (n = 2).