(A & C) RT-qPCR analysis of tax/rex mRNA levels from untreated (blue) or stimuli treated (red) FS cells. FS cells were incubated with PMA (A) or vorinostat (C) over the times indicated, prior to harvest for RNA analysis. Data are normalized to Ct values for ASL and divided by the value obtained for the zero time point. Arrows indicate time points of harvest for nuclear run-on assays. (B & D) Nuclear run-on assay for measuring newly transcribed mRNA. Nuclei were isolated at the indicated times [15 minute and 5 hours of PMA treatment (B), 5 hours and 9 hours of vorinostat treatment (D)], and incubated with biotinylated uridine, and biotinylated transcripts were isolated. Transcripts corresponding to full length HTLV-1 and ASL were detected by RT-qPCR, and the data normalized to ASL. Results represent the fold change over the value obtained for time zero. Average fold change with standard deviation (error bars) from triplicate inductions are shown.