Fig. 9

Analysis of gD mutants for the ability to restrict the release of infectious HIV-1. 293 cells were co-transfected with vectors expressing gD, gD[ΔTMCT], gD[ΔCT], and gD _[6−259], gB (negative control), or UL47 (negative control) and a plasmid pNL4-3. At 48 h, the culture supernatants were collected and the levels of infectious virus released into the culture supernatants was determined using TZM-bl cell assays and levels of p24 determined using p24 kits. a Schematic diagram of the gD mutants analyzed. b Results of restriction analysis of gD and different gD mutants. c The levels of p24 protein released from cells from the experiments in (b). d The expression of each gD protein and β-actin (left two panels), gD proteins released into the culture medium (middle panel) and from UL47 and β-actin (right panels) from a representative restriction assay using immunoblots. The experiments were performed at least four times and statistical differences with the pcDNA3.1(+)/HIV-1 control evaluated using a two-tailed Student’s t-test, with p < 0.01 (filled triangle) considered significant