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Fig. 1 | Retrovirology

Fig. 1

From: Effect of induced dNTP pool imbalance on HIV-1 reverse transcription in macrophages

Fig. 1

Effect of single dNTP elevation imbalances on HIV-1 infection in macrophages. All experiments were conducted in human primary monocyte-derived macrophages prepared from four healthy donors. dA and dT treatments were at 2.5 mM and dG treatment was at 1 mM, due to solubility limits. a dNTP levels were measured after macrophages were treated with a single dN treatment. After 12 h single dN treatment, the dNTP levels in these cells were determined by the RT-based dNTP assay [8]. “+” indicates the dNTPs of the corresponding treated dNs, and “x” indicates untreated dNTPs. NT No dN treatment. b HIV-1 vector transduction efficiency was measured after macrophages were pretreated with dNs for 4 h, and then transduced with an equal amount of HIV-GFP vector. The transduced cells were collected after 6 days, and the percent of the GFP+ cells was determined by FACS. The ratio of the vector transduction efficiency at each single dN treatment was normalized to the transduction efficiency with no dN treatment, which gave ~ 10% transduction efficiency. See Additional file 1: Table S2 for Raw %GFP numbers. c Reverse transcription kinetics was determined by measuring the copy numbers of HIV-1 2-LTR circle DNAs in the cellular genomic DNAs isolated from the treated macrophages. d p24 antigen levels were measured by ELISA. Macrophages were treated with single dNs for 4 h followed by dual tropic HIV-1 89.6 infection. Supernatant was collected 6 days after infection. Supernatant was used for a HIV-1 p24 ELISA. The measurement ratios were normalized with the no treatment (NT) conditions, and marked at the top of each readout. The data in this figure are the mean of three independent experiments and error bars represent the standard deviation from the mean. *p-value < 0.05

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