Effects of AG490 on expression of KSHV-encoded genes and cytokines in PEL cells.
(A) AG490 minimally affects expression of antiapoptotic proteins encoded by KSHV. Cells were treated with 100 μM AG490 or DMSO for 24 hours. RT-PCR analysis was performed with the use of DNase-treated total RNA (0.25 μg for each reaction) for KSHV viral FLICE-inhibitory protein (vFLIP) (orf K13/orf 71), v-cyclin (orf 72), latency-associated nuclear antigen (LANA) (orf 73), v-survivin (orf K7), and v-IL-6 (orf K2). KSHV−/EBV+ Daudi cells were used as a negative control. The quality of RNA was confirmed by parallel RT-PCR amplification for GAPDH. (B) Regulation of growth factor secretion by AG490. Cells were treated with 100 μM AG490 or 0.1% DMSO for 24 hours, and specific ELISAs were performed on culture supernatants. Mean (± SD) protein levels per 106 live cells are shown from 3 independent experiments: SDs lower than 5% of the mean values are not visible in the bar graphs.
