FIGURE 9.

Role of RNF4 on EBV lytic development. A, P3HR1 cells were infected by the lentivirus that contains RNF4 shRNA (shRNF4) or control shRNA (Ct-shRNA) under the selection of puromycin. Thereafter, cells were treated with sodium butyrate and TPA (SB/TPA) for 48 h, and proteins in the lysate were examined by immunoblotting using anti-Rta, anti-EA-D, anti-RNF4, and anti-α-tubulin antibodies. B, moreover, cells that were harvested from A were lysed, and EBV lytic DNA replication assay was assayed by qPCR. The amount of EBV DNA was normalized with the amount of actin DNA that was determined in the same assay. C, the lentiviral-transduced P3HR1 cells were treated with TPA and sodium butyrate for 5 days. EBV DNA from viral particles that were released into the culture medium was determined by qPCR after the DNA extraction. The copy number of EBV genome was calculated by using maxi-EBV that had been isolated from E. coli as a standard.