Fig. 5: RB but not E2F1 expression level is the decisive factor for viral genome replication.

a, b siRNA-mediated knockdown of one of E2F1,3 and 4 was performed with siPOOL technology in T24 cells infected with ADWT or XVir-N-31 (MOI 50). a Viral genome replication was analyzed at 24 hpi. n = 4. b Quantification of viral particles was performed by a hexon titer test and presented as infectious units per milliliter (IFU/ml). n = 2, mean ± SE. c Viral replication was analyzed in T24 cells transfected with siE2F1 pool and treated with 500 nM Palbociclib for 24 h, and infected with XVir-N-31 (MOI 50). d siRNA-mediated knockdown of E2F1 was performed with siPOOL technology in T24 cells and infected with ADWT or XVir-N-31 (MOI 50). Protein levels were detected by immunoblotting at indicated time points for E2F1, RB, and E1A proteins. GAPDH was used as a reference protein. e SK-N-MC cells were transfected with siRNAs against E2F1 or RB or RBL1 (p107) or RBL2 (p130) and infected with the XVir-N-31 (MOI 20). Viral replication was analyzed at 48 hpi. n = 3. f RB-negative T24shRB1 cells and scrambled control T24shCtrl cells were treated with Palbociclib (1 µM) and infected with MOI 50 of the indicated viruses. Viral genome replication was analyzed at 24 hpi. g RB-negative Saos-2 cells were transfected with indicated plasmids and infected with adenovirus ADWT (MOI 20). Viral genome replication was analyzed at 24 hpi. n = 3. All viral genome replication analyses were performed by qPCR to amplify viral fiber DNA. Data are represented as relative fiber DNA at indicated time points compared to fiber DNA at 4 hpi as the baseline (mean ± SD). SD standard deviation, SE standard error, MOI multiplicity of infection, hpi hours post-infection. The statistical significance was determined by a two-sided Student’s t-test; n: number of biologically independent samples. Source data are provided as a Source Data file.