Virus infections and inhibitors
1 x 106 HFFF-TERTs were plated in a 25
cm2 flask. After 24 h, the medium was changed to DMEM
lacking FBS but with 4 μg/ml dexamethasone, as this approach has been
shown to improve infection efficiency (Tanaka et al., 1984). After 24 h,
the medium was changed to DMEM containing the requisite volume of HCMV
strain Merlin stock to achieve MOI 5. Cells were gently rocked (5 rpm)
for 2 h, and then the medium was changed to DMEM/FBS/PS. MG132 (Sigma)
at 10 μM or bortezomib (Sigma) at a range of concentrations was added to
the cell culture 12 h prior to sample collection. For 12 hpi
experiments, inhibitors were added to the initial viral mixture used for
infection, which was replaced with drug-containing fresh DMEM after the
2 h of incubation.