FIG 2.
Raltegravir can inhibit DNA replication of HSV-1(F). CV1 cells were infected with virus at an MOI of 5 PFU per cell. After virus adsorption and washing, cells were maintained in medium containing DMSO alone or with 200 μM raltegravir. (A) The cells were lysed, and total cellular DNA was extracted, digested with BamHI, electrophoretically separated on 0.8% agarose gels, and transferred to nitrocellulose. (B) The gels were then probed with radiolabeled BamHI P fragment, which is specific for sequences at genomic ends (P fragments), and internal junction fragments (S-P fragment). (C) Comparison of the DNA copies of the viral genome of HSV-1(F) with DMSO treatment and raltegravir treatment using real-time quantitative PCR. Target primers for UL42 and reference primers for 18S rRNA were used to quantify viral DNA. The relative amount of amplicon DNA was calculated by subtraction of the CT value of target genes and 18S rRNA (control) gene in the raltegravir-treated samples. This difference was then substracted from the same calculation derived from the DMSO-treated samples by the 2−ΔΔCT method. qPCR was performed in triplicate, and data are shown as the mean ± SD of three independent experiments. ***, P < 0.001.