MuHV-4 gH Figure 4. gL2 virions show altered glycoprotein and capsid distributions and gB conformation changes. A. NMuMG cells were exposed to wild-type or gL2 virions (2 h, 4uC, 3 p.f.u./cell) then washed 63 in PBS and either fixed immediately (4uC) or first incubated (2 h, 37uC) to allow virion endocytosis (37uC). The cells were then permeabilized and stained for LAMP-1 plus MuHV-4 capsid or glycoprotein antigens as shown. Nuclei were counterstained with DAPI. Colocalization appears as yellow in the merged images. Equivalent data were obtained in 3 further experiments. B. MCCD epithelial cells were incubated with a range of different gL2 or gL+ viruses (2 h, 4uC, 3 p.f.u./cell), then washed 63 with PBS and incubated further (2 h, 37uC) to allow virion endocytosis. The cells were then stained for gp150, an abundant virion glycoprotein, with mAb LS-B11 (IgG1) and for ORF65 capsid with mAb MG-12B8 (IgG2a). Nuclei were counterstained with DAPI. doi:10.1371/journal.pone.0002811.g004 PLoS ONE | www.plosone.org 5 July 2008 | Volume 3 | Issue 7 | e2811