Figure 2
From: Studies on membrane topology, N-glycosylation and functionality of SARS-CoV membrane protein
N-glycosylation of coronviral M. A, N-terminal amino acid sequences of M of selected coronaviruses were aligned and depicted using MacVector program. B, M and the glycosylation-deficient mutant MN3Q of HCoV-NL63 were in vitro translated and analyzed as described above. (Plasmid expressing HCoV-NL63 M protein was kindly provided by M. Müller, Bonn). C, Subconfluent Huh7 cells were transfected with plasmid encoding the glycosylation-deficient M (MN4QN-FLAG) using FuGENE according to the manufacturer's protocol. At 24 h p.t., cells were fixed, permeabilized and incubated with a polyclonal α-FLAG antibody and a rhodamine-coupled secondary antibody. ERGIC was stained using a monoclonal α-ERGIC primary antibody and a FITC-coupled secondary antibody. The merged image is shown on the right.