Fig. 5

Identification of inhibitors of HIV-1 Gag expression from the GSK PKIS library. a Structures of 1H3 and 2E3. Indicated are the EC₅₀ and CC₅₀ values of the compounds as determined from assays using HeLa C7 cells; b HeLa C7 cells were incubated with compounds at increasing compound concentration and HIV-1 gene expression induced with Dox (4.5 µM) for 24 h. 1% DMSO treated cells grown with or without Dox served as positive and negative controls, respectively. Dose response on HIV-1 gene expression was measured relative to intracellular GagGFP levels in DMSO & Dox-treated samples. Effects of compounds on cell viability were assessed using alamarBlue assay across n > 3 independent assays. c, d CEM-HIV* cells were treated with compounds 1H3 (200 nM) or 2E3 (100 nM) and HIV-1 gene expression induced with Dox and prostratin. 1% DMSO treated cells grown with or without Dox and prostratin served as positive and negative controls, respectively. Cells were harvested for HIV-1 protein and RNA analyses after 24 h of induction. c Shown are representative western blots showing the effect of compounds on HIV-1 Gag, Env, and Tat levels. d Quantification of viral TAR, US, SS, and MS RNA levels in cells treated with compounds relative to induced DMSO control. The positions of the primers in the proviral construct are shown in Fig. 2A. Viral mRNA levels were normalized to PUM1 for TAR RNA and ß-actin for US, SS, and MS RNA. Mean mRNA levels were expressed relative to Dox and Prostratin induced DMSO control. Data are indicated as mean ± SEM, n = 4 independent experiments, *p ≤ 0.05, **p ≤ 0.01, and ***p ≤ 0.001