Fig. 8.

Evaluation of viral replication efficiency in stably transduced HFFs expressing doxycycline-inducible anti-cyclin H shRNAs. HFF WT (A) or stably transduced HFFs, under conditional dox-inducible KD, termed HFF sp3 (C) and HFF sh53 (E) were seeded in 12-well plates before shRNA was induced by the addition of 500 ng/ml dox for 5 d. Next, dox-induced and non-induced HFFs were infected with HCMV AD169 at MOI of 0.25 and were harvested at indicated time points for standard SDS-PAGE and Wb analysis using the indicated antibodies. Additionally, dox-induced and non-induced HFFs were infected with HCMV AD169 at MOI of 0.001 (B, D, F). Viral supernatants were collected at the indicated time points and viral genome equivalents were determined by qPCR. Mean values ± SD of two independent biological replicates, each measured in technical duplicates, are shown. Statistical analysis was performed using an ordinary two-way ANOVA and post-hoc Sidak correction: ***, p < 0.001; ****, p < 0.0001.