Analysis of cell death in HIV-1-infected HLAC. HLACs were infected with 6 ng p24 of wt HIV-1, Δnef HIV-1 and the two Nef variants LLAA and Δ12–39 in quadruplicates. On day 8 p.i., cells were stained for 7AAD, CD3, CD8 and intracellular p24. Results from one single infection are shown. (A) Flow cytometry analysis of the CD3/CD8 stain of live T lymphocytes. (B) Flow cytometry analysis of CD3+/CD8- lymphocytes (based on the gating in A) for intracellular p24 and 7AAD for the detection of productively HIV-1 infected and dead CD4+ T cells, respectively, (C) Percentage of total productively infected (p24+) CD4+ lymphocytes in the HLAC infection shown in A. (D) Percentage of dead (7AAD+) and infected (p24+) CD4+ T lymphocytes in the HLAC infection shown in A. (E) Percentage of dead (7AAD+) uninfected (p24-) CD4+ T lymphocytes in the HLAC infection shown in A. Mock designates the percentage of dead CD4+ T lymphocytes in the mock-infected culture.