Fig. 2
Activation of latent integration-competent HIV-1 and unintegrated HIV-1 with a panel of latency reversing agents (LRA). a Experimental design. Raltegravir was added on days 0, 3 and 7 and not removed until sorting. b The responses of latently infected cells to various LRAs under No RAL and +RAL conditions, expressed as the percentage of GFP+ cells 1 day after stimulation. Data from one experiment representative of >3 independent experiments with cells from different donors is shown. Each condition was tested in triplicate. Additional file 1: Figure S5 shows an independent experiment (Experiment 2 in Fig. 3c, d, g, h) employing an expanded panel of LRA. Similar results were obtained with the Integrase D116N mutant (not shown). c The percentage of GFP+ cells generated in the No RAL vs. the +RAL cultures for each LRA in two independent experiments from b (Expt. 1) and in Additional file 1: Fig. S5 (Expt. 2). Each symbol represents one LRA from Fig. 2b (Experiment 1) and S5 (Experiment 2). d Virus production from No RAL vs. +RAL cultures for both experiments. e The percentage of GFP+ cells generated by various LRAs vs. virus release into the culture medium to compare virus production per GFP+ cell. The dynamic range is shown for the number of GFP+ cells and for virus production as fold induction of the maximum over the minimum value. Data are from Expt. 1 and is representative of 3 additional independent experiments. f GFP Mean fluorescence intensity (MFI) of the GFP+ cells vs. virus production for No RAL and +RAL cells in Expt. 1. Similar results were obtained from Expt. 2 (not shown). The dynamic ranges are shown as in e. g Relationship between the strength of the LRA in inducing virus production (X axis) and the +RAL output expressed as a percent of the No RAL output. +RAL output reached 66 % of No RAL output for Bryostatin+ SAHA. h Relationship between the strength of the LRA in inducing virus production (X axis) and the +RAL output per GFP+ cell expressed as a percent of the No RAL output per GFP+ cell. c–h All p ≤ 0.001