FIG 8.

Analysis of EBV lytic replication and gene expression induced by mutant BZLF1 in BZLF1-KO EBV BAC-harboring B cells and epithelial cells. (A to E) Wild-type and BZLF1-KO EBV BAC-harboring BJAB cells were infected with GFP-tagged wild-type BZLF1 or BZLF1Δ207-210-expressing lentiviruses (MOI = 10). (A) At 48 h postinfection, BZLF1-KO EBV BAC-harboring BJAB cells were collected and subjected to immunofluorescence (IF) staining with anti-BZLF1 antibody. (B and C) At 48 h postinfection, wild-type and BZLF1-KO EBV BAC-harboring BJAB cells were collected, whole-cell extracts were analyzed by Western blotting as indicated (B), and viral mRNAs were extracted, reverse transcribed, and analyzed for expression by reverse transcription-quantitative PCR (RT-qPCR) (C). (D and E) The cell pellets and supernatants were collected at different time points after lentiviral infection, and then intracellular viral genomic DNA (D) and extracellular virion DNA (E) were extracted and analyzed by real-time PCR. *, P < 0.05. (F and G) Wild-type and BZLF1-KO EBV-BAC-harboring HNE-1 cells were seeded in a 6-well plate and transfected with 2 μg control vector or GFP-BZLF1- or GFP-Δ207-210-expressing plasmids. The expression levels of viral proteins were determined by Western blotting 48 h posttransfection (F), and intracellular viral genomic DNA was extracted and analyzed 72 h after transfection (G). *, P < 0.05.