Fig. 6.

The gB(275Y) variant accelerates viral entry. (A and B) MRC-5 fibroblasts were infected by AD169 gB(275Y) or gB(275D) (MOI 3) and harvested at the indicated times postinfection. Viral protein levels were determined by immunoblot analysis; β-actin was used as a loading control. (C) MRC-5 fibroblasts were pretreated for 30 min with 100 µM EIPA or vehicle. Two hours postinfection with AD169 gB variants (MOI 3), cells were fixed and the viral tegument protein pp65 was detected by indirect immunofluorescence. Nuclei were stained with DAPI. (Scale bars, 20 µm.) Images representative of 3 independent experiments are shown. (D) MRC-5 cells were pretreated and infected as above. Two hours postinfection EIPA was removed and fresh medium was added. Cells were harvested at the indicated times and IE1 expression was analyzed by immunoblot. The data shown are representative of 3 independent experiments.