Fig 7.

gL-deficient virions show altered glycoprotein distribution during entry. (A) MDBK or EBL cells were exposed to WT BAC, gL STOP, or gL STOP Rev BoHV-4 strains (3 h, 4°C, 10 PFU/cell) and then washed in PBS and either fixed immediately (0 h postbinding) or incubated first (3 h, 37°C) to allow virion endocytosis (3 h postbinding). The cells were then permeabilized and stained for BoHV-4 glycoprotein antigen (MAb 35, anti-gB). Nuclei were counterstained with To-Pro-3. Equivalent data were obtained in three further experiments. (B) BoHV-4 virions were bound to MDBK cells (3 h, 4°C, 10 PFU/cell) with or without bafilomycin (5 nM). The cells were then washed in PBS to remove unbound virions and further incubated with or without drug (3 h, 37°C) to allow virion endocytosis. The cells were then fixed, permeabilized, and stained for gB plus LAMP-1.