Effect of alsterpaullone in PBMC infection. A) Phytohemagglutinin (PHA) and IL-2 activated PBMCs were kept in culture for 2 days prior to infection. Approximately 5 × 106 PBMCs were infected with pNL4-3 (MOI:1). Alsterpaullone treatment (0.01-5.0 μM) was used (only once) immediately after the addition of the fresh medium. Samples were collected every sixth day and stored at -20°C for further analysis (RT assay). Both PHA and IL-2 were added to media every 3 days. Viral supernatants (10 μl) were incubated in a 96-well plate with reverse transcriptase (RT) reaction mixture, incubated overnight at 37°C, spotted, washed, dried, and then counted using a Betaplate counter. B) Cells were also counted (~100/date) for viability using trypan blue staining method. C) Approximately 5 × 106 activated PBMCs were infected with primary HIV-1 strain (THA/92/00NSI), and then treated after viral adsorption (12 hrs) with either mock (DMSO) or cyc202 (0.01 μM) or alsterpaullone (0.01 μM) or with the combination of both drugs. Samples were collected every six days (0, 6, 12, 18 and 24 days) and stored at -20°C or p24 assay.