Figure 1

HIV-1 latency assay. A: Schematic of the HIV-1 latency assay. SupT1 T cells are infected with HIV-1 for 4 hours, free virus is washed away, and the fusion inhibitor T1249 is added to prevent new infections. Infected cultures are split 24 hours after infection into a mock and anti-latency drug treated culture. Cells are harvested 24 hours after treatment, stained for intracellular CA-p24 and analyzed by FACS. The fold activation (as viral latency marker) is the ratio of CA-p24 positive cells in the drug versus mock treated sample. B: Representative FACS analysis. Live cells are gated using the Forward/Sideward scatter (FSC/SSC) and scored for CA-p24 positivity in the RD1 channel. C: Latency assay: percentages of CA-p24 positive cells in control (mock treated), TNFα treated, Vorinostat treated and DMSO treated (mock for Vorinostat treated) cultures. The results presented are the average values of two independently produced virus stocks, which were both used in two independent infections. Significant difference (*) was determined with the student T-test (Graphpad Prism). D: The fold activation (percentage CA-p24 positive cells in drug induced culture versus mock culture).