Fig. 5

Specific cellular signaling pathways with associated host cellular factors are involved in reactivation of latent HIV-1 in ACH-2 cells. a ACH-2 cells were pretreated with specific inhibitors IΚK2 inhibitor V (50 µM), FK506 (10 µM), Cyclosporin A (10 µM), SP600125 (50 µM), SB203580 (50 µM), U0126 (50 µM), AZD6244 (10 µM), WP1066 (5 µM), Rottlerin (50 µM), or vehicle control (DMSO), and 4 h later activated with SAHA (1 µM, white bars), prostratin (1 µM, grey bars) or TNF-α (0.1 ng/ml, bars with diagonal line upwards). HIV reactivation was estimated at 12 h following reactivation, by intracellular p24 Gag staining by flow cytometry. For comparison of results across samples from multiple experiments, HIV-1 reactivation observed in vehicle control pretreatment was considered as 100 % and the background (no reactivating agent) as 0 %. Error bars represent standard deviation (N = 3), *p < 0.05. b At the end of 16 h following addition of inhibitors, the live cells were evaluated by Trypan blue staining. The percentage of viable cells was calculated by subtracting the dead cells from total cells divided by total cell count. c The amount of p24-Gag positive cells was estimated by intracellular p24-Gag staining and flow cytometry. Error bars represent standard deviation (N = 3), *p < 0.05