FIG. 7.

Membrane-targeted Nef activates HIV-1 LTR in cooperation with Tat in Jurkat T cells. (A) The CD8-Nef chimera was described previously (4). The truncated CD8 molecule (CD8ET) that encodes only extracellular and transmembrane domains was used as a control. CD8-Nef (1 μg) or CD8-ET (1 μg) was cotransfected with or without Tat (0.05 μg) together with the wild-type LTR (LTRwt)-SEAP reporter vector (0.2 μg) into Jurkat TAg cells. The SEAP activity was measured 48 h after transfection. (B) The cooperative effect of the CD8-Nef requires the κB and TAR elements. The transient cotransfection was performed by using CD8-Nef and Tat together with the LTRwt-SEAP, LTRΔκB-SEAP, or LTRΔTAR-SEAP in Jurkat cells. The data are representative of two independent experiments.