FIG 7.

DNA replication and ChIP assay. 293T cells were transfected with plasmids for OriP control lacking the EBNA1 gene (pHEBO) or OriP vectors in pREP10 expressing WT EBNA1 or the R481E or D591E mutant. (A) Western blot for EBNA1 and the loading control, actin. (B) Southern blot analysis of plasmid DNA replication assay for EcoRV-linearized (left) and EcoRV- and DpnI-digested (right) DNA probed with ³²P-labeled OriP DNA. (C) Quantification of Southern blot DNA replication assay for WT and mutant EBNA1 shown in panel B. (D) ChIP assay with antibodies to IgG, EBNA1, or MCM3 and assayed by quantitative PCR (qPCR) at AMP, DS, and FR regions of the pREP10 plasmid.