HIV-1 Latency Reactivation Assay

J-Lat C11 cells^{24 (link),26 (link)} (established in our lab) and A10.6 cells^{29 (link),54 (link)} (obtained from NIH AIDS Reagent Program) harboring latent, transcriptionally competent HIV-1 provirus that encodes GFP as an indicator of viral activation were cultured in RPMI1640 medium (Gorning) supplemented with 10% fetal bovine serum (FBS) (Gibco), 100 U/mL penicillin and 100 μg/ml streptomycin (Gibco) in a 37 °C incubator containing 5% CO₂. For in vitro reactivation experiments, C11 cells or A10.6 cells were stimulated with RVX-208 (Selleckchem), PFI-1 (Selleckchem), JQ1 (Selleckchem), SAHA (Sigma-Aldrich), prostratin (Sigma-Aldrich) or TNF-α (Sigma-Aldrich) respectively and subjected to determine GFP expression using flow cytometry (BD Calibur).