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The K111R mutant reduces unwinding of the replication origin. C33A cells were transfected with FLAG-E1, FLAG-E2, or PFLORI31. Unwinding at origin DNA was analyzed by FAIRE assay using HPV31 LCR primers LCR3 and LCR4 (A). (B) ChIP for RPA protein at the LCR used anti-RPA70 antibody and LCR4 primers. *, P < 0.05 by one-way ANOVA compared to the wild type in the absence of E1; #, P < 0.05 by one-way ANOVA compared to wild-type E2 plus E1.
