Figure 2
HT ligand-based FACS analysis of HIV-1 Lai-Halo -infected cells and HEK-293 cells expressing a HT-p65 fusion protein. A, B, C FACS analysis of Jurkat cells infected with HIV-1Lai-Halo with a FACSAria cytometer equipped with a 488- and 561-nm lasers.. A. Oregon Green ligand staining. (blue: uninfected cells; green: virus-infected cells); B. TMR ligand staining (blue: uninfected, unstained cells; red: uninfected, stained cells; green: infected, stained cells). C. FACS analysis of fixed Jurkat cells stained with TMR ligand comparing with unfixed cells (blue: uninfected cells; red: virus-infected cells fixed with PFA; green: virus-infected, unfixed cells). Data were acquired with DiVa 6.1 software 5 days postinfection and analyzed off-line with the FlowJo program (Treestar, Ashland, OR). D. Microscopic analysis of Jurkat cells infected with HIV-1Lai-Halo and stained with the Oregon Green HT ligand. E. FACS analysis of HEK-293 cells expressing the HT-p65 fusion protein and stained with different doses of Alexa 488 HT ligand (red: unstained cells; blue, green and orange: 0.25, 0.5, and 1.0 μM, respectively). F. Control HEK-293 cells unstained and stained with Oregon Green HT-ligand (red and blue, respectively) and cells transfected with p65-expressing plasmid and stained with 0.5 and 1.0 μM Oregon Green HT-ligand (green and orange, respectively).