FIG 7.

PML and capsid assembly by EBV. (A) P3HR1 cells were untreated (Latent) or treated with TPA and sodium butyrate (T-S). After treatment, cells were transfected with pCMV-BDLF1 or pCMV-BFRF3 or cotransfected with these two plasmids to express BDLF1 and BFRF3. (B) Cells were infected with lentivirus expressing control shRNA (Ctrl-sh) or PML-specific shRNA (PML-sh) for 5 days and then treated with TPA and sodium butyrate or left untreated. Three days after lytic induction, the quantity of capsids produced by the cells was analyzed via ELISA. EBV capsids were captured by anti-BFRF3 antibody and detected using rabbit anti-BDLF1 antibody and goat HRP-conjugated anti-rabbit immunoglobulin antibodies. (C) P3HR1 cells (1 × 10⁵) were transfected with control shRNA and PML-specific shRNA. Cells were treated with TPA and sodium butyrate and cultured in 30 ml medium for 5 days. EBV particles were collected from the culture medium by centrifugation. The EBV genome in the viral particles was measured by real-time qPCR. (D) The PML, Rta, VCA, BORF1, BDLF1, and α-tubulin expressed by the cells were analyzed via immunoblotting 3 days after lytic induction.