FIG. 5.

GCV partially prevents changes in the abundance of eIF4F components and the composition of eIF4F complexes, but it has no effect on 4E-BP1 phosphorylation or activation of ERK1/2 and p38. (A) Quiescent NHDFs were mock infected (M) or infected with HCMV AD169 (MOI = 5) in the presence or absence of ganciclovir (150 μM). Total protein was harvested at 72 h postinfection, fractionated by SDS-PAGE, and analyzed by immunoblotting using the indicated antisera. (B) As in panel A, except that detergent extracts were prepared at 72 h postinfection, and proteins bound to 7-methyl GTP-Sepharose 4B were fractionated by SDS-PAGE, immunoblotted, and visualized with the indicated antibodies. (C) NHDFs were either mock infected (M), infected with gradient purified, UV-inactivated HCMV AD169 (MOI = 5) (UV), or infected with wild-type untreated virus (WT). Total protein was isolated at 12 h postinfection, fractionated by isoelectric focusing (for eIF4E) or SDS-PAGE, and analyzed by immunoblotting with the indicated antisera. At 2 h postinfection, cells were fixed and stained with a monoclonal antibody against the pp65 tegument protein to demonstrate that the UV-treated virus was still capable of entering cells.