Fig. 9

BRD4 mediates HIV D4-A7 RNA splicing. HEK293T cells were co-transfected with the pLTR.gp140/EGFP.Rev∆38/DsRed splicing reporter, 20 ng of pRev and 100 ng of pTat101 (AD8) without (wild-type, WT) or with specific mutations (K28A, K50A, K50/51A, K71A and R53A) then treated with DMSO, 1 μM JQ1 (+) or JQ1 (−). Cells were harvested at 48 h, EGFP and DsRed expression was quantified by flow cytometry. a The flow cytometry gating strategy used to identify  % EGFP and  % DsRed positive cells in a representative result from n = 3 independent experiments, each conducted in triplicate. b The mean percentage of spliced product DsRed/(DsRed + EGFP) from the 3 independent experiments is shown. Comparisons of each condition to JQ1 (+) were made using Friedman nonparametric test. Only statistically significant comparisons are shown *p < 0.05; **p < 0.01