Figure 2

INI1 coactivates Tat-mediated HIV-1 transcription. a. P4.2 cells (2 × 10⁴ cells per well) were cotransfected with 100 ng of pHIV-1-LTR-Luc [25], 100 ng of pcDNA3-Tat101-FLAG [41], and/or 200 ng of pCGN-INI1 [7]. Luciferase assay was performed 24 h later. All transfections utilized equal total amounts of plasmid DNA owing to the addition of empty vector into the transfection mixture. Results were obtained through three independent transfections. Relative stimulation of luciferase activity (fold) is shown. b. 100 ng of pcDNA3-Tat101-FLAG, and/or 200 ng of pCGN-INI1 were cotransfected into P4.2 cells in triplicate. Beta-galactosidase activity was measured 24 h later. c. P4.2 cells were cotransfected with 100 ng of pHIV-1-LTR-Luc, 50 ng of pH2F Tat (Tat86) or pH2Tat (Tat72) [26], and/or 200 ng of pCGN-INI1 and performed luciferase assays 24 h later. d. 100 ng of pHIV-1-LTR-Luc, and/or 100 ng of pcDNA3-Tat101-FLAG were cotransfected into INI1 knockdown P4.2 cells (INI1i) or P4.2 cells transduced with a control (C) lentiviral vector and luciferase assays were performed 24 h later. e. INI1 deficient MON cells (2 × 10⁴ cells per well) were cotransfected with 100 ng of pHIV-1-LTR-Luc, 50 ng of pcDNA3-Tat101-FLAG, and/or 200 ng of pCGN-INI1 and luciferase assays performed 24 h later.