Fig 2: CDC6 expression in ccRCC tissues and adjacent normal renal tissues. IHC staining showed low CDC6 expression in normal renal tissues (A, B) and low (C, D), medium (E, F) and high (G, H) ccRCC tissues.

Fig 3: Associations between CDC6 and PPI, MSI, Neoantigen, TMB in ccRCC. (A) Associations between CDC6 and MSI; (B) Associations between CDC6 and Neoantigen; (C) Associations between CDC6 and TMB; (D) PPI network.

Fig 4: Gene and protein expression analysis of replication regulating genes CDC6, CDT1, and GMNN. HSkM cells were differentiated for 24 h and RNA was isolated at the indicated time points and used for RT-qPCR (A). QuantiTect Primer Assays for CDT1, CDC6, and GMNN were used as test genes and compared to TBP and GAPDH housekeeping genes. RT-qPCRs were done in two replicates and standard deviation is indicated as a vertical line. HSkM cells were differentiated for 24 h and protein was isolated at the indicated time points and used for Western blot analysis (B). Primary antibodies against CDT1, CDC6, and GMNN were probed in separate blots and each blot was sequentially probed with ß-actin as loading control. Protein sizes were indicated and revealed expected protein sizes for all tested antibodies.

Fig 5: 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 HAN?RNA2.7 (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 HAN?RNA2.7 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 HAN?RNA2.7 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 HAN?RNA2.7 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.