Fig. 1

Establishment of HIV latency in macrophage-like THP-1 cell line. (A) Screening of latency-reversing agents in latently infected THP-1. Initial screening of LRAs in a mixture of latently infected non-clonal THP-1 cells. (B) Reactivation of HIV in latently infected non-clonal dTHP-1 cells. uTHP-1 cells were infected with the HIV-1 reporter virus (HIVGKO) for 4 days and sorted for cells expressing mKO2 (latently infected cells). Cells were transformed into macrophage-like cells by differentiation with 10 ng/ml PMA for 48 hours followed by reactivation with JQ1 (1000 nM) for 48 hours. Cells were then fixed and observed under a fluorescent microscope. Cells with the reactivated virus show csGFP. Percentages show mKO2 + cells that also became csGFP + after reactivation (csGFP+, mKO2+). (C) PCR of DNA extracted from expanded single clones with primers covering HIV-1 5’ LTR and envelope to further screen for clones with full-length viral DNA. Clones with full-length single viral copies of integrated viral DNA are shown with asterisks (*). These clones were selected for further characterization. Data are means and error bars indicate ± SEM (n = 3). *p < 0.5, p < 0.1, **, p < 0.01; ***, p < 0.001, ns, not significant, Student’s t test