Fig 8.

Exogenous expression of BPLF1 deubiquitinase or p65 knockdown restores viral DNA replication of the BPLF1-deficient virus. (A) The BZLF1 expression plasmid (0.5 μg/well) was transfected into 293EBVΔ cells using an electroporator, and 4 h after the initial transfection, the cells were further transfected with wild-type or enzyme-dead BPLF1 expression plasmids (0.5 μg/well) using Lipofectamine 2000. At 48 h after the initial transfection, cells were washed with PBS (−), and total DNA was extracted. qrt-PCR analysis was performed with the same method as described for Fig. 7B. cDNAs were prepared from the mRNAs extracted in parallel with the total DNAs. RT-PCR data from one representative experiment are shown below the graph. (B) p65-targeted or control siRNA (0.2 μg/well) was cotransfected with the BZLF1 expression plasmid (0.2 μg/well) into 293-EBVΔ cells using an electroporator and cultured for 48 h and processed similarly to the method described for panel A. Data are expressed as fold increase in comparison to untransfected cells and means ± SD of the results of 3 biological replicates. **, P < 0.001; *, P < 0.01.