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. 2009 Jun 3;83(16):8090–8098. doi: 10.1128/JVI.02379-08

FIG. 6.

FIG. 6.

Camptothecin inhibits Zta assembly of replisome at OriLyt. (A) ZKO-293 cells were transfected with Zta expression vector, followed by treatment with 1 μM camptothecin (CTN) (+) or vehicle control (−). Cells were then assayed by ChIP for Zta binding to viral OriLyt or to the cellular actin locus. (B) ZKO-293 cells transfected with Zta and then treated with (+) or without (−) camptothecin were assayed by ChIP with antibodies to Topo I (αTopoI) or control IgG. ChIP DNA was assayed at EBV OriLyt (left bars) or the control region in cellular actin (right bars). (C) ZKO-293 cells were transfected with FLAG-Zta or control FLAG expression vector and then subjected to FLAG immunopurification (IP). FLAG affinity-purified proteins were assayed by Western blotting (immunoblotting [IB]) for the cellular RecQL1 protein (left) or by colloidal blue staining of the affinity-purified proteins (right). FLAG-Zta is indicated by the arrow. (D) A ChIP assay was used to monitor RecQL1 binding to OriLyt in vivo in ZKO-293 cells transfected with FLAG-Zta followed by the addition of camptothecin (+) or vehicle control (−). ChIP DNA was assayed at the EBV OriLyt or cellular actin locus and quantified as change over control IgG.