Figure 8.

Inhibiting eIF4E phosphorylation significantly reduces HSV-1 replication and protein synthesis in quiescent cells infected at low input doses of virus. (A) Growth-arrested NHDF cells were either mock infected (M) or infected with HSV-1 (MOI = 10; 15 h postinfection) in the presence and absence of the indicated inhibitors. eIF4E and 4E-BP1 in whole-cell lysates were detected by IEF or SDS-PAGE followed by immunoblotting with the appropriate antisera. (B) Primary human fibroblasts (NHDF cells) were growth arrested by serum starvation and infected at low multiplicity in the presence or absence of the p38 inhibitor SB203580, the mnk inhibitor CGP57380, or rapamycin. After 5 d, cell-free lysates were prepared by freeze-thawing and titered in Vero cells. (C) NHDF cells growth arrested and treated as in A were either mock infected or infected with wild-type (WT) HSV-1 Patton at the indicated multiplicities. After a 1-h pulse with ³⁵S amino acids at 14 h postinfection, cell-free lysates were prepared and either immunoprecipitated with anti-HSV-1 antisera (MOI = 0.01) followed by SDS-PAGE, or directly analyzed by SDS-PAGE (MOI = 1 and 10).