Figure 3

Effect of SMAPP1 on HIV-1 gene expression in chronically HIV-1 infected T cell lines and primary cells. a Chronically HIV-1 infected T cell line ACH-2 was pre-treated with the cocktail of antiretrovirals (lamivudine/emtricitabine, tenofovir and indinavir each in 10 µM concentrations) for 7 days, then the cells were washed and cultured in regular medium with indicated concentrations of SMAPP1 or SAHA. The cells were harvested at 24 and 48 h after the treatment, total RNA was isolated and subjected to quantitative RT-PCR analysis with the primers specific for HIV-1 gag. Results are shown as a mean of three independent experiments ±SD. Asterisk shows p value ≤0.05; double asterisk shows p value ≤0.01 between control cells and cells treated with SAHA. b The same cells were treated with a lower concentration of SMAPP1 or SAHA. Quantitative RT-PCR analysis of total RNA with oligo-dT (RT reaction) and gag-specific primers (qPCR) was performed in 24 h post-treatment. Results are shown as a mean of three independent measurements ±SD. Asterisk shows p value ≤0.01 between the control cells and cells treated with 3 µM SMAPP1. c, d Statistical analysis of the effect of SMAPP1 and bryostatin 1 on HIV-1 transcription in low-productive infected PBMCs. The PBMCs from healthy donors were activated with IL-2, infected with HIV-1 subtype B strains NL4-3 (c) or subtype C 1084i (d) (20 ng of p24 per 10 × 10⁶ cells) by spinoculation and after 8 days cultivation in medium with IL-2 were cultured for 15 days with IL-7 to transfer T cells to quiescent phase. The cultures were treated with SMAPP1 or bryostatin 1 and then cultured for 48 h. Asterisk shows p value ≤0.01 between the control cells and cells treated with bryostatin 1.