Figure 4. Effect of nocodazole on KSHV virion titers.

BCBL-1 cells were treated with increasing concentrations (0.5, 5.0, 10.0 µM) of the microtubule depolymerizing drug nocodazole and induced with TPA. For nocadazole washout experiments, cells were treated with 5 µM of the drug, washed thoroughly to ensure complete removal of the drug, and then induced with TPA. Following induction, extracellular KSHV virions were collected and concentrated 100-fold. Virus stocks were treated with Turbo DNase I and viral DNAs extracted. KSHV genomic DNA concentration was estimated by a real-time PCR along with external standards of known concentrations of the viral DNA using primers against the ORF73 gene and expressed as viral DNA copy numbers per milliliter (ml) of the supernatant.