Figure 8.

HSV-1 induced clustering of receptor and identification of virions containing vesicles. (A) Nectin-1 colocalizes with early stage vesicles in HSV-1 infected cells. CHO cells transfected with a plasmid expressing nectin-1-EGFP were incubated with Texas red-transferrin conjugate (1:250) for 10 min and then either (a–c) mock infected or (d–f) infected with HSV-1 (50 PFU/cell) for 30 min, fixed and examined by confocal microscopy. Nectin-1 expression is seen in green (a and d) and the early stage vesicles appear red (b and e). In mock-infected cells, merger of green and red shows no colocalization (c), but in infected cells there is colocalization (yellow) (f); (B) HSV-1 colocalizes in early stage vesicles. As indicated, nectin-1-CHO or CHO-WT (Control) cells were infected with HSV-1 (K26GFP) (green) for the time points indicated and then treated with citrate buffer (pH 3) and fixed. It was followed by staining with either anti-EEA1 or anti-TfR (red) to identify the vesicles. By sequential CM the z stacks show colocalization of virions with early stage vesicles. Little colocalization with recycling to late endosomes was observed.