FIG. 11.
Accumulation of viral mRNAs and proteins in MEF lines infected with HSV-1(F). Two sets of MEFs in T25 flasks were exposed to 0.1 PFU/cell of HSV-1(F) and harvested at 8 or 12 h. One set of cells was lysed in RIPA buffer, and the whole-cell proteins were extracted and subjected to electrophoresis in a denaturing gel, transferred to a nitrocellulose membrane, and reacted with antibody to ICP4, ICP0, ICP27, TK, or US11. The parallel set of cells was lysed in 1 ml TRIzol reagent, and total RNA was extracted, 10 μg of which was resolved by electrophoresis in a 1% denaturing formaldehyde agarose gel and transferred onto a nylon membrane. The membrane was then probed with 32P-labeled DNA probes specific for mRNA of ICP4, ICP0, or ICP27. The top gel shows electrophoretically separated total RNA stained with ethidium bromide.