CD4+ T cells from 6 virally suppressed HIV-1–infected individuals were activated with anti-CD3/CD28 antibodies for 4 days in the presence of HIV-1–suppressing agents (10 μM) and ART (1 μM tenofovir and 10 μM enfuvirtide) to allow cellular proliferation without new rounds of infection. Cells were plated at limiting dilution (200,000 cells per well) to calculate the frequency of cells harboring inducible HIV-1. After 2 days allowing washout of the HIV-1–suppressing agents, cells were stimulated with PMA/ionomycin to induce HIV-1 RNA expression. The use of inducible HIV-1 RNA assay by measurement of supernatant HIV-1 RNA allows a wide dynamic range to calculate the frequency of cells harboring inducible HIV-1. P values were calculated by Friedman’s nonparametric ANOVA test (2-tailed) with uncorrected Dunn’s test for comparison between each treatment and DMSO control. DM, DMSO; Fi, filgotinib; Ru, ruxolitinib; Sp, spironolactone; MA, mycophenolic acid.