FIG 5.

NF-κB activation and BART RNA and miRNA expression in EBV-infected epithelial cell lines. (A) Immunoblot analysis of nuclear and cytoplasmic fractions from the cell lines C666-1, HeLa-Bx1, and AGS-Bx1, using specific antibodies. Antibodies against lamin A+C were used to verify preparation of nuclear and cytoplasmic fractions. (B) EBV copy numbers in the cell lines C666-1, HeLa-Bx1, and AGS-Bx1 were determined by amplifying a fragment of the BamH-W region using qPCR. (C) Relative levels of BART miRNA in the EBV-infected epithelial cell lines C666-1, HeLa-Bx1, and AGS-Bx1 were determined by real-time PCR. BART miRNA expression was normalized to that of miR-Hsa-16. (D) Relative levels of the alternatively spliced BART RNAs BARF0, RPMS1, RPMS1A, and A73 in the EBV-infected epithelial cell lines C666-1, HeLa-Bx1, and AGS-Bx1 were determined by real-time PCR. BART RNA expression was normalized to that of GAPDH. Real-time PCR experiments were performed in triplicate, and the results are the averages and standard errors of the means from three independent RNA extractions. P values were calculated using a two-tailed Student's t test (*, P < 0.05; ns, not significant).