Fig. 3

JQ1 induces HIV spliced RNA accumulation in the absence and presence of Tat. a Schematic of unspliced and spliced mRNAs produced following HEK293T cell transfection with the pLTR.gp140/EGFP.Rev∆38/DsRed splicing reporter. The different sets of primers-probe used for HIV RNA quantification are displayed as coloured arrows for primers, and linked coloured spots for the probe. The env set detects unspliced mRNA (green), rev set detects spliced mRNA (red) and the viral set detects all viral mRNAs including the co-transfected pRev^NL4.3 mRNA (brown). b–d HEK293T cells transfected in the absence or presence of 100 ng of pTat101 (AD8)-Flag expression plasmid were treated for 24 h with JQ1 (1 μM) or DMSO diluent control (n = 4). Cells were then harvested and EGFP (unspliced) and DsRed (spliced) protein expression measured using flow cytometry (Fig. S2), while HIV unspliced (US), spliced (D4-A7) and all viral RNA expression levels (copies/μl) were quantified by droplet digital PCR (ddPCR) (b–d). The ratio of US (c) and spliced (d) over all viral RNAs is displayed as fold-change (FC) over DMSO. Comparisons of each condition to DMSO were made using a paired T test. Only statistically significant comparisons are shown *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. The black lines represent the mean ± SEM