The eEF1A inhibitor didemnin B blocks HIV-1 reverse transcription.

<p>(<b>A</b>) Jurkat cells transfected with pCMV-Gluc 2 were treated post-transfection with media containing didemnin B (Did B) or cycloheximide (CHX) at concentrations as indicated. The levels of secreted <i>Gaussia</i> luciferase in the culture supernatants were measured 8 h post-treatment. (<b>B</b> and <b>C</b>) Jurkat cells were incubated with subnanomolar concentrations of Did B and CHX as indicated and then infected with HIV-1 at 4°C for 2 h and 37°C for 4 h. The total cytoplasmic DNA was collected and the levels of HIV-1 early (<b>B</b>) and late (<b>C</b>) reverse transcription products were measured by qPCR. (<b>D</b>) Reverse transcription efficiencies were calculated as the ratio of late to early viral DNA, expressed as a percentage. (<b>E</b>) The levels of HIV-1 RNA in cells treated with didemnin B (Did B) and cycloheximide (CHX). Jurkat cells were incubated with 100 nM nevirapine and concentrations of Did B and CHX as indicated at 37°C for 2 h followed by HIV-1 infection. The HIV-1 was incubated with cells at 4°C for 2 h and then at 37°C for 4 h. Heat inactivated virus was used as a negative control. The HIV-1 RNA was extracted from the cells and detected by RT-qPCR. A direct qPCR without RT from the RNA samples was performed to monitor for DNA contamination. All columns represent a mean value and standard deviation from at least three independent experiments. ** = <i>p</i> < 0.05.