Figure 3
From: Studies on membrane topology, N-glycosylation and functionality of SARS-CoV membrane protein
Characterization of recombinant SARS-CoV expressing a glycosylation-deficient M. A, Vero cells were infected with wild type recombinant SARS-CoV (wt rec SARS-CoV) or the mutant MN4Q recombinant SARS-CoV (MN4Q rec SARS-CoV) respectively with an moi of 3 pfu per cell. Cell lysates were harvested and subjected to Western blot analysis. Virus inactivation was achieved by treatment with 1% SDS and boiling of the samples. Proteins were separated by SDS-PAGE and blotted onto PVDF membranes. To detect viral structural proteins the membrane was incubated with a serum of an immunized rabbit or with the human monoclonal antibody S30. B, For electron microscopic analysis, Vero cells were infected with a MOI of 1 with wt rec SARS-CoV or MN4Q rec SARS-CoV, respectively. At 24 h p.i., viral particles were fixed, negatively stained and subjected to electron microscopic analysis.