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. 2011 Jul;85(13):6127–6135. doi: 10.1128/JVI.00258-11

Fig. 1.

Fig. 1.

The majority of viral genome DNA is localized inside BMRF1-rich structures. (A) Lytic replication-induced Tet-BZLF1/B95-8 cells were fixed, stained with anti-BMRF1 (green) and anti-BALF2 (red) antibodies, and observed by laser scanning confocal microscopy to locate viral replication compartments. The right panel is a merged image. (B) Lytic replication-induced Tet-BZLF1/B95-8 cells were fixed in 4% paraformaldehyde and 70% ethanol. After digestion with RNase, they were treated with 50% formamide, air dried, and immediately hybridized with a mixture containing the EBV BAC DNA probe labeled with DIG nick translation mix. Specimens were stained with fluorescein isothiocyanate (FITC)-conjugated anti-DIG sheep (green) and anti-BMRF1 mouse (red) monoclonal antibodies. The right panel is a merged image. (C) Newly synthesized DNAs were labeled by incubation with 10 μM CldU added directly to the culture medium of lytic replication-induced Tet-BZLF1/B95-8 cells for 10 min at 24 h postinduction in the presence (b) or absence (a) of PAA (400 μg/ml). Specimens were stained with anti-BALF2 (green), anti-CldU (yellow), and anti-BMRF1 (red) antibodies.