J-Lat 9.2 or 10.6 cells were seeded in 96 well plates at 200,000 cells/well and co-treated with a pro-inflammatory cytokine (10 ng/mL TNFα) or an LRA (10 μM prostratin, 0.1 μM panobinostat, 10 μM JQ1, or 10 mM HMBA) in the presence of test agents at defined concentrations or 0.1% DMSO vehicle control for 24 hours. Cells were then examined for GFP expression by flow cytometry. In a subset of experiments, cells were fixed and permeabilized using the Cytofix/Cytoperm fixation kit (BD Biosciences; Mississauga, ON, Canada), stained for intracellular p24Gag (KC57-RD1 antibody, Beckman Coulter; Mississauga, ON, Canada), and examined for p24Gag expression by flow cytometry.
OM-10.1 cells were seeded as above and cotreated with 1 ng/mL TNFα and test agents or 0.1% DMSO control for 24 hours. Cells were fixed, permeabilized, stained for p24Gag as above, and examined for p24Gag expression by flow cytometry.
OM-10.1 cells were seeded as above and cotreated with 1 ng/mL TNFα and test agents or 0.1% DMSO control for 24 hours. Cells were fixed, permeabilized, stained for p24Gag as above, and examined for p24Gag expression by flow cytometry.