Fig. 2.

Cyclin upregulation upon HCMV infection in TEV-1 cells and HFFs. TEV-1 cells (A) or HFFs (D) were seeded in 12-well plates and infected 1 d later with HCMV Merlin at MOI of 0.5. Cells were harvested at indicated time points in duplicates originating from different wells (see duplicate samples indicated by odd- and even-numbered lanes) before cells were lysed and prepared for standard SDS-PAGE and Wb analysis using the indicated antibodies. Two independent SDS-PAGEs/Wbs were used for densitometry measurements in quadruplicate using ImageJ. Cyclin H signal intensity was normalized to β-actin and set in relation to mock-infected controls for infected TEV-1 cells (B) and infected HFFs (E). TEV-1 cells (C) or HFFs (F) were infected in 12-well plates with HCMV Merlin at MOI of 0.5 to assess cyclin H mRNA levels. Optionally, cells were treated with 1 µM of MBV or equal amounts of DMSO. Cells were harvested at the indicated time point, total RNA was extracted and cyclin H mRNA levels were determined by a one-step RT-qPCR using cyclin H-specific primers and probe. The values obtained were normalized to the mock-infected controls. Statistical analysis was performed using an ordinary one-way ANOVA test and post-hoc Tukey correction: ns, not significant; *, p<0.05; **, p<0.01; ***, p<0.001; ****, p<0.0001.