Figure 4

(A) Percentage of GFP-positive K562 cells expressing NC or MLST8 sgRNAs 21–24 h after PMA stimulation with or without simultaneous Torin1 treatment. Data are represented as mean +/− SD of four independent experiments.

(B) Detection of selected phosphorylated proteins in CD4 T cells from four independent donors using PathScan analysis. CD4 T cells were treated with either 0.01% DMSO or incubated for 30 minutes with 25µL of αCD3/αCD28 activating beads with DMSO, 250nM pp242, 97.7nM Torin1 or 10nM rapamycin. The amount of phosphorylated proteins is scaled internally to each donor.

(C–E) Bcl-2 transduced latently infected cells were either unstimulated (yellow) or stimulated with CD3/CD28 antibodies (2.5 µg/ml CD3 and 0.65 µg/ml CD28 (orange); 10 µg/ml CD3 and 0.65µg/ml CD28 (red)) for 48 h to reactivate HIV in the presence of increasing concentrations of pp242 (C), Torin1 (D) and rapamycin (E). Reactivation of HIV was assessed by measuring GFP by flow cytometry, and the percentages of reactivation were calculated for each batch of latently infected cells by maximum activation with PMA and Ionomycin (top panels) (Yang et al., 2009). Percentage of live cells in each sample is shown (bottom panels). Data are represented as mean +/− SD.

See also Figure S2