Figure 4
From: Selective Targeting of Virus Replication by Proton Pump Inhibitors
EBV replication is tenatoprazole-sensitive. Latent EBV was reactivated in CLIX-FZ cells by addition of doxycycline in the presence of DMSO (black bars) or DMSO plus tenatoprazole (20 µM or 40 µM, grey bars); non-treated cells are shown as white bars. After 72 hr, cells and conditioned media were harvested. Panel A, extracellular EBV DNA measured in pellet fraction of conditioned media. Panel B, intracellular EBV DNA measured in lysed cells. EBV DNA was determined by quantitative-PCR (qPCR) for the EBV BALF5 gene. EBV DNA copy number from each preparation was normalized to the DMSO-treated control. Panel C, metabolic activity of cells in the presence of tenatoprazole. The cells were grown for 24 hr in media containing the indicated concentrations of doxycycline and tenatoprazole followed by assessment of metabolic activity assessed using the WST-1 assay. Panel D, fractionation of intracellular DNA. CLIX-FZ cells were treated with doxycycline plus DMSO (D+) or doxycycline plus tenatoprazole (D+T), harvested 72 hr later, and separated into nuclear (black bars) and cytosolic (white bars) fractions. DNA was extracted from each fraction and the relative number of EBV genomes from each fraction was determined by qPCR using primers directed towards the EBV BALF5 gene. The number of genomes recovered was significantly different as judged by the Students t-test, two-tailed. Error bars equal 1 SD.
