Fig 3.

AS601245 prevents HIV-1 reactivation without inhibiting NF-κB. (A) Latently HIV-1-infected CA5 T cells were left untreated or pretreated with AS601245 for 1 h. Then the cells were left either untreated or stimulated with PMA to trigger HIV-1 reactivation as indicated. Levels of HIV-1 reactivation were determined by flow cytometric analysis for GFP expression 24 h after PMA stimulation. (B) From the cultures depicted in panel A, samples were taken at the indicated time points (0 to 60 min) and used to determine the level of induced NF-κB activity in the presence or absence of AS601245 by using TransAM assays. (C) Experiments similar to those described in the legend to panel B using HRF as the HIV-1-reactivating stimulus. (D) CA5 T cells were stimulated with TNF-α, and NF-κB activity was followed over a 500-min period to determine any long-term effects of AS601245 on TNF-α-induced NF-κB activity oscillation. (E) Inhibitory effect of the NF-κB inhibitor aloisine A and the CDK inhibitor roscovitine on HIV-1 reactivation in CA5 T cells. (F) Effect of the PI3 kinase inhibitor Ly294002, AKT inhibitor VIII, or the GSK inhibitor TDZD-8 on HIV-1 reactivation in CA5 T cells. All experiments were done as dose matrix experiments, and the depicted conditions represent maximum achievable inhibition at a cell viability of >50%.