FIG. 3.

Rate-zonal centrifugation of HSV-1 capsids isolated from the nuclear fraction and analyzed for the presence of tegument proteins. Capsids were obtained from the nuclear fraction of HSV-1-infected Vero cells harvested at 15 h after infection and separated on a 20–50% linear sucrose gradient following rate-zonal centrifugation. 0.5 ml fractions were collected from the bottom of the tube. The proteins within each fraction were precipitated with 10% TCA, separated by SDS-PAGE followed by Western blot analysis using primary antibodies to specific tegument proteins followed by the addition of the secondary anti-rabbit or anti-mouse antibody conjugated to horseradish peroxidase. Western blot analysis was done using antibodies to capsid proteins VP5 (the major capsid polypeptide) and VP21/VP22a (scaffold proteins), tegument proteins VP1/2 and UL37, and the DNA binding protein UL42.