Fig. 5

Gag-fusion viruses display defects in Gag processing and are less able to saturate TRIM5α. A: Immunoblot of WT LAI, 8.91 LAI, Gag-LUC and Gag-GFP virus particles (2 × 10¹¹ genomes) detecting HIV-1 p24, firefly luciferase or GFP and a schematic of Gag cleavage products. MA: matrix, CA: capsid, SP1: spacer peptide 1, NC: nucleocapsid, SP2: spacer peptide 2 and location of the GFP/luciferase gene insertion. B: Abrogation-of-restriction assay in FRhK4 cells expressing restrictive rhesus TRIM5. FRhK4 cells were co-transduced with a fixed dose of WT LAI.GFP (5 × 10⁷ genomes/ml) and increasing doses of the WT/Gag-LUC chimeric viruses carrying a luciferase-expressing genome (0.0005–1 U RT/ml). Rescue of GFP infectivity was assessed by flow cytometry at 48 h. Data are singlet % GFP values and two repeats of the experiment are shown. Statistical analyses were performed using 2-way ANOVA with multiple comparisons. * P < 0.05, n.s. non-significant