FIG. 6.

Presence of DNA DSBs in the viral genome synthesized in the replication compartments. TUNEL staining, performed as described in Materials and Methods, was used to detect DSBs in viral replication compartments. Tet-BZLF1/B95-8 cells were cultured in the presence of 2 μg of doxycycline/ml and harvested at 24 h p.i. Cells were treated with 0.5% Triton X-100-mCSK buffer, fixed with methanol, and coimmunostained with anti-BMRF1 (blue) antibody and anti-RPA32 Ser-33 (red) or anti-Rad51 (red) antibodies and with fluorescein-dUTP in the presence of TdT (green) as described in Materials and Methods. (Right) Merged images. (IR) for a TUNEL-staining positive control, Tet-BZLF1/B95-8 cells were exposed to 10 Gy of gamma irradiation and harvested 4 h posttreatment. Cells were stained with fluorescein-dUTP in the presence of TdT.