Skip to main content
. 2004 Oct;78(20):11084–11096. doi: 10.1128/JVI.78.20.11084-11096.2004

FIG. 5.

FIG. 5.

Relationship between Nef modifications and changes in Nef-induced apoptosis. (A) The changes in Nef-induced apoptosis are not the results of instability of the resultant Nef protein. Virions were transcomplemented with mutant Nef proteins, and the resultant viruses were analyzed for infectivity and the ability of the Nef protein to enhance infectivity. Bar 1 represents the infectivity of the wild-type HIV-1 virus in a MAGI assay; bar 2 represents the infectivity of a virus with a deletion of Nef; bars 3, 4, and 5 represent the infectivities of viruses with a deletion of Nef that were transcomplemented with normal Nef (bar 3), NefΔM1 (bar 4), or NefΔM1M2 (bar 5). The error bars show the standard errors of the measurements, and the results are a compilation of at least three independent experiments. (B) The changes in Nef-induced apoptosis are not the results of variations in Nef protein expression. A Western blot analysis with 40 μl of HEK 293 conditioned medium was performed by using an anti-Nef antibody. Lane 1: untransfected HEK 293 cell conditioned medium; lane 2: HIV Nef protein (1 μg); lane 3: HIV-1 Nef-transfected HEK 293 cell conditioned medium; lane 4: HIV-1 NefΔM1-transfected HEK 293 cell conditioned medium; lane 5: HIV-1 NefΔM1M2-transfected HEK 293 cell conditioned medium. (C) Deletion of apoptosis motifs in the Nef protein eliminates Nef-induced apoptosis. Jurkat cell cultures were treated with conditioned cell supernatants from HEK 293 cells expressing various Nef variants to analyze the effect of the changes in Nef on Nef-induced apoptosis. Bar 1 represents the results for Jurkat cells treated with medium (UT); bar 2 represents treatment with Nef protein (N; 4.3 nM); bar 3 represents treatment with conditioned medium from untransfected HEK 293 cells; bar 4 represents treatment with 40 μl of conditioned medium from pNL4-3 KFS-transfected HEK 293 cells; bar 5 represents treatment with 40 μl of conditioned medium from pNefΔM1-transfected HEK 293 cells; and bar 6 represents treatment with 40 μl of conditioned medium from pNefΔM1M2-transfected HEK 293 cells. The error bars show the standard errors of the measurements, and the results are a compilation of at least three independent experiments.