Figure 5

Stable TRN-SR2 depletion inhibits single round infection by HIV-1 WT and N74D CA mutant virus. (A) shTR3, shTR4 and shSCR HeLaP4 cells were challenged using 3 dilutions of VSV-G pseudotyped HIV-1 NL4-3 (WT) or (B) HIV-1 NL4-3 N74D CA mutant (N74D) luciferase reporter viruses. Two days post infection Fluc activity was measured and normalized to total protein amounts. Graphs show the mean values of Fluc light units per μg protein (Fluc LU/μg protein) ± SD of one representative experiment out of two performed in triplicate. (C) Same as in (A), but 2 dilutions of multiple-round viruses HIV-1 NL4-3 (WT) or (D) HIV-1 NL4-3 N74D CA mutant (N74D) carrying the HIV-1 envelope were used in the presence of 5 μM ritonavir to ensure single round infection. Infectivity was measured by β-gal activity 72 hours post infection. The arrows indicate the relative inhibition of infectivity in the shTR3 and shTR4 HeLaP4 cells compared to shSCR control cells.