Reverse Transcriptase

The Nucleotide analog reverse-transcriptase inhibitor Tenofovir disoproxil fumarate was added to TZM-bl cells. Cells were inoculated with 0.05 ng mock-exposed and semen-exposed HIV, and 0.5 ng HIV as infectivity-matched control. Infection rates were measured 3 days post infection by measuring β-galactosidase or or 4 days post infection by measuring luciferase activities. The left panels show the mean enzyme activities ± standard deviation derived from triplicate infections. RLU/s: relative light units per second. Middle panels show normalized infection rates in which reporter enzyme activities obtained from infected cells in the absence of inhibitor were set at 100%. The right panels depict the calculated IC50 values. The number above the bar represents the fold-change in the IC50 derived from 0.05 ng semen-exposed relative to 0.05 or 0.5 ng mock-exposed virus infection. Ns, no statistically significant difference; **** p<0.0001; *** p<0.001 (unpaired t-test).

Human PBMCs containing indicated concentrations of Tenofovir disoproxil fumarate were inoculated with 5 ng mock-exposed or semen-exposed R5-HIV-luciferase, or 50 ng R5-HIV-luciferase as infectivity matched control. Infection rates were determined 3 days post inoculation.

Cells were inoculated with 0.05 ng mock-exposed and semen-exposed HIV, and 0.5 ng HIV as infectivity-matched control. Infection rates were measured 3 d post infection by measuring β-galactosidase. Left: the mean enzyme activities. Middle: normalized infection rates in which reporter enzyme activities obtained from infected cells in the absence of nevirapine were set at 100%. Right: IC50 values.

The indicated liver cancer cell lines—Hep-3B, Huh-7 and PLC were treated with or without 5 types anti-hepatitis B virus drugs (C) with the concentration of 100 μM, and M1 virus (MOI = 10) for 72 hours. Following 72 hours, cell viabilities were determined by MTT assay (mean ± SD). N.S. Not significant.

1D 19F NMR spectra of RT181tfmF and several RT181tfmF mutants in the absence (black) and presence of NVP (pink), EFV (green), ETR (blue) and RPV (orange). (A) Superposition of the 19F spectra of apo-RT181tfmF and the 19F spectra of RT181tfmF in the presence of each NNRTI, (B) Superposition of the 19F spectra of apo-RT181tfmF-V108I and the 19F spectra of RT181tfmF in the presence of each NNRTI, (C) Superposition of the 19F spectra of apo-RT181tfmF-K103N and the 19F spectra of RT181tfmF-K103N in the presence of each NNRTI, (D) Superposition of the 19F spectra of apo-RT181tfmF-E138K(p51) and the 19F spectra of RT181tfmF-E138K(p51) in the presence of each NNRTI. The 19F spectra in the presence of each NNRTI at 1:1 and 1:5 molar ratios are shown in light and dark colors, respectively. Chemical formulae for each inhibitor are depicted in the individual panels.

The Nucleotide analog reverse-transcriptase inhibitor Tenofovir disoproxil fumarate was added to TZM-bl cells. Cells were inoculated with 0.05 ng mock-exposed and semen-exposed HIV, and 0.5 ng HIV as infectivity-matched control. Infection rates were measured 3 days post infection by measuring β-galactosidase or or 4 days post infection by measuring luciferase activities. The left panels show the mean enzyme activities ± standard deviation derived from triplicate infections. RLU/s: relative light units per second. Middle panels show normalized infection rates in which reporter enzyme activities obtained from infected cells in the absence of inhibitor were set at 100%. The right panels depict the calculated IC50 values. The number above the bar represents the fold-change in the IC50 derived from 0.05 ng semen-exposed relative to 0.05 or 0.5 ng mock-exposed virus infection. Ns, no statistically significant difference; **** p<0.0001; *** p<0.001 (unpaired t-test).

Human PBMCs containing indicated concentrations of Tenofovir disoproxil fumarate were inoculated with 5 ng mock-exposed or semen-exposed R5-HIV-luciferase, or 50 ng R5-HIV-luciferase as infectivity matched control. Infection rates were determined 3 days post inoculation.

Cells were inoculated with 0.05 ng mock-exposed and semen-exposed HIV, and 0.5 ng HIV as infectivity-matched control. Infection rates were measured 3 d post infection by measuring β-galactosidase. Left: the mean enzyme activities. Middle: normalized infection rates in which reporter enzyme activities obtained from infected cells in the absence of nevirapine were set at 100%. Right: IC50 values.

The indicated liver cancer cell lines—Hep-3B, Huh-7 and PLC were treated with or without 5 types anti-hepatitis B virus drugs (C) with the concentration of 100 μM, and M1 virus (MOI = 10) for 72 hours. Following 72 hours, cell viabilities were determined by MTT assay (mean ± SD). N.S. Not significant.

1D 19F NMR spectra of RT181tfmF and several RT181tfmF mutants in the absence (black) and presence of NVP (pink), EFV (green), ETR (blue) and RPV (orange). (A) Superposition of the 19F spectra of apo-RT181tfmF and the 19F spectra of RT181tfmF in the presence of each NNRTI, (B) Superposition of the 19F spectra of apo-RT181tfmF-V108I and the 19F spectra of RT181tfmF in the presence of each NNRTI, (C) Superposition of the 19F spectra of apo-RT181tfmF-K103N and the 19F spectra of RT181tfmF-K103N in the presence of each NNRTI, (D) Superposition of the 19F spectra of apo-RT181tfmF-E138K(p51) and the 19F spectra of RT181tfmF-E138K(p51) in the presence of each NNRTI. The 19F spectra in the presence of each NNRTI at 1:1 and 1:5 molar ratios are shown in light and dark colors, respectively. Chemical formulae for each inhibitor are depicted in the individual panels.