Fig. 6.

Specific mutations within Tat reduce HIV D4-A7 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 without (WT) or with specific mutations (K28A, K50A, K50/51A, K71A and R53A). Cells were harvested at 48 h and DsRed expression was quantified by flow cytometry. a A representative example of the gating strategy used to identify  % DsRed positive cells that represent spliced product. b The proportion of spliced product [DsRed/(DsRed + EGFP) × 100] relative to WT Tat is shown from n = 5 independent experiments, each conducted in triplicate. Comparisons of each condition to DMSO were made using a Paired T test. Only statistically significant comparisons are shown **p < 0.01; ***p < 0.001. The black lines represent the mean ± SEM