Fig. 9.

Viral replication efficiency in transduced HFF cell populations carrying an expression module of inducible anti-cyclin H shRNA sp3. (A) Non-transduced HFFs WT or transduced HFFs carrying the dox-inducible anti-cyclin H shRNA sp3 module were seeded in 12-well plates and shRNA was induced by the addition of 500 ng/ml dox for 5 d. Thereafter, 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. KD efficiency of cyclin H was quantitatively determined by densitometric evaluation. Dox-induced and non-induced HFF sp3 were infected with HCMV Merlin WT (B) or mutant Merlin expressing GCV resistance-conferring A594V variant of pUL97 (Schütz et al., 2022) (C) at MOI of 0.001. 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 < 0.0001.