Human MDM were plated in 24-well plates and then infected with HIV-1ADA for 24 hours before incubation with ONC201 or ONC201 isomer at 3, 10, or 30 μM for five days. A) Cell lysates were collected and subsequently subjected to SDS-PAGE and immunoblotting for the detection of phospho-FOXO3a, total FOXO3a, and p24. Actin was used as the loading control. Densitometric quantifications of phospho-FOXO3a in MDM were presented as a ratio to total FOXO3a and normalized as fold changes to the vehicle control DMSO group. B) Soluble TRAIL concentrations after ONC201 or ONC201 isomer treatment were determined from cell culture supernatants using ELISA. Data were normalized with total cellular protein concentrations. Results shown are from representative experiments performed with three different donors. ANOVA analysis: * denotes p <0.05, ** denotes p < 0.01, compared to the vehicle control DMSO group.