Fig. 3.

Regulation of host cell cycle by HCMV RNA2.7. A Block of host cells entry into S phase by RNA2.7. HELF cells were infected with HAN or HANΔRNA2.7 (MOI ​= ​1.0). DRB was added 4 ​h before infection with a final concentration of 200 ​nmol/L and was used as a positive control. Cells were stained and analyzed by flow cytometry. Percentages of cells in the G0/G1 phases of the cell cycle were calculated and presented as mean ​± ​SEM. B Model of pre-RC formation for cellular DNA replication. Components involved in pre-RC formation were up-regulated in HANΔRNA2.7 infected cells and were facilitated cellular DNA replication. Gene that mRNA fold change more than 2 is indicated with red color. Gene that mRNA fold change less than −2 is indicated with green color. C Effects of RNA2.7 on genes facilitating pre-RC formation. Transcriptions of MCM2, MCM4, MCM5, Cdt1 and Cdc6 were increased in HANΔRNA2.7 infected cells, and could be decreased by inhibiting Pol II S2 phosphorylation. Data are presented as mean ​± ​SEM. D Effects of RNA2.7 on expressions of Cdt1 and Cdc6. Protein levels of Cdt1 and Cdc6 were increased in HANΔRNA2.7 infected cells, and could be decreased by inhibiting Pol II S2 phosphorylation. Data are presented as mean ​± ​SEM. Statistical analysis was performed by Student's t-test. ∗P ​< ​0.05; ∗∗P ​< ​0.01; ∗∗∗P ​< ​0.0001. DRB, 5,6-dichloro-1-b-D-ribofuranosyl benzimidazole; pre-RC, pre-replication complex.