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. 2016 Oct 4;6:34532. doi: 10.1038/srep34532

Figure 1. The 5′-LTR from HIV-1 B′ subtype demonstrates the strongest capacity for driving gene expression.

Figure 1

(a) Schematic construction of pGL3 containing LTR-consensus-driven luciferase reporter. (b,c) Assay for LTR-driven gene expression upon stimulation. HIV-1 LTR-luciferase-reporter plasmids and pRenilla-luc-TK were co-transfected into HEK293T cells for 24 h and stimulated with TNF-α (b) or PMA/Ionomycin (c) for additional 24 h. The cells were harvested, and luciferase activity was measured and calculated by using dual luciferase reporter assay. (d) Schematic construction of pHIV-eGFP containing LTR consensus. (e) Infection of Jurkat CD4+ T cells. Jurkat CD4+ T cells were infected with pHIV-eGFP/VSV-G (1 ng p24gag amount viruses for 106 cells) for 24 h, with or without TNF-α (2 ng/ml) stimulation, and viral infection was determined by measuring GFP expression with flow cytometry, and the mean fluorescence intensity (MFI) was calculated. *P < 0.05, **P < 0.01 and ***P < 0.001 are considered significant difference in an unpaired t-test.