Fig 2.
Impact of UL83 mutagenesis on pp71 expression levels and DNA replication kinetics. (A) Immunoblot analyses of lysates from cells infected with the indicated viruses at 125 genomes per cell and collected at the indicated times after infection. Lysates from 1 × 105 cells were applied to each lane on SDS-polyacrylamide gels. Antibodies directed against pp71 or β-actin were used for detection. (B) Quantification of the pp71 bands from panel A, using the Odyssey analysis system. Signals from individual bands were normalized to β-actin in each case (RFI, relative fluorescence intensity). (C) Quantification of viral DNA replication at low infectious doses. Cells were infected with either 0.4 genome copies per cell (top graphs) or 0.04 genome copies per cell (bottom graphs). Infected cells were collected at the indicated time points of infection. Cellular DNA was purified and analyzed by quantitative DNA PCR, using HCMV-specific primers.