HA-tagged WT E2F1, E2F1-KK, and E2F1 R109K plasmids have been described previously (11). HA-tagged E2F1 L132E and R166H constructs were generated from WT HA-E2F1 using a site-directed mutagenesis kit (Stratagene). Flag-tagged SENP7 V5 was generated by subcloning from an open reading frame shuttle clone (0CAAo5051G027D; Source BioScience) using primers targeting the start and stop codons (flanked with NotI and SalI restriction sites, respectively). The polymerase chain reaction (PCR) product was purified using a PCR purification kit (Qiagen) and digested with the required enzymes (Promega) for 1 hour. The digested DNA was gel-purified using a gel extraction kit (Qiagen) and ligated into the p3xFlag-CMV-7.1 vector (Sigma-Aldrich). Plasmid transfections were performed for 48 hours using the GeneJuice transfection reagent (Novagen), as per the manufacturer’s instructions. RNA interference was performed with 25 nM siRNA for 72 hours using the Oligofectamine transfection reagent (Invitrogen), as per the manufacturer’s instructions. Sequences for siRNA are as follows: nontargeting control, 5′-AGCUGACCCUGAAGUUCUU-3′; E2F1, 5′-CUCCUCGCAGAUCGUCAUCUU-3′; p100/TSN, 5′-AAGGAGCGAUCUGCUAGCUAC-3′; SENP7, 5′-GAAGUAAGACAGUAGAUGA-3′.

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