FIG 8.

Effect of 5LO inhibition on KSHV gene expression and protein levels in PEL cells. (A to H) BCBL-1 or BC-3 cells were treated with 5LO chemical inhibitors (1 μM MK866 and 1 μM zileuton) or their respective solvents or transduced with sh-C or sh-5LO, and RNA was prepared. Viral transcripts (vCyclin, LANA-2, LANA-1, and vFLIP) were quantitated by real time-RT PCR as described before (19). Percent inhibition was calculated by considering viral gene expression in solvent-treated or sh-C-transduced cells to be 100%. Each point represents the average ± SD from three independent experiments. The statistical analysis was carried out using a two-tailed Student's t test. *, P < 0.05; **, P < 0.01. Red arrows facing down, inhibition. (I and J) Effect of 5LO inhibition on viral proteins LANA-1, vCyclin, and LANA-2. Cell lysates prepared from BCBL-1 or BC-3 cells treated with 5LO chemical inhibitors (1 μM MK866 and 1 μM zileuton) or their respective solvents or transduced with sh-C or sh-5LO were Western blotted for LANA-1, vCyclin, and LANA-2, stripped, and immunoblotted for α-tubulin. Representative blots from three independent experiments are shown.