Extended Data Fig. 7: TNFα can be used instead of Tax to augment transcription from vectors comprising a CMV promoter.
(a) Left; schematic of pcDNA plasmid encoding EGFP under a CMV promoter in the sense orientation. Middle; bar graph representing % EGFP expressing HEK293T cells 48 hours after transfection with suboptimal levels of plasmid in the presence or not of co-transfected plasmid encoding Tax, soluble TNFα, or PMA. Right; bar graph shows relative mean fluorescence intensity (MFI) of EGFP under the different experimental conditions (EGFP encoding plasmid alone is set to 100%). (b) Schematic of sense lentiviral vector encoding EGFP and produced in absence or presence of TNFα (c) Left; titer measurement expressed as Transducing Units (TU) per ml for three independent experiments. Middle; transduction of Jurkat cells with decreasing volumes of lentivirus vector supernatant to evaluate percentage EGFP expression by flow cytometric analysis on day 5. Bar graph represents the mean of three independent experiments. Right; representative histograms of Jurkat cells transduced with 30 µl sense and antisense lentivirus vector supernatant.
