Nef Alleles from All Major HIV-1 Clades Activate Src-Family Kinases and Enhance HIV-1 Replication in an Inhibitor-Sensitive Manner
Figure 10
Inhibition of HIV-1 Nef chimera-mediated activation of SFKs by 4-amino DFP analogs.
CEM-T4 cells were infected with wild-type HIV-1 NL4-3 (WT), a Nef-defective mutant (ΔNef), or the indicated Nef chimeras in the absence or presence of 3 µM DFP-4A, DFP-4AP, DFP-4AB, or the carrier solvent (DMSO) alone as a control (Con). SFK proteins were immunoprecipitated from infected cell lysates followed by immunoblotting with a phosphospecific antibody against the activation loop phosphotyrosine residue common to all SFKs (pY418). Control blots were performed on cell lysates for HIV-1 Gag proteins (p55, p40, and p24), Nef, as well as actin as a loading control. Results from uninfected cells are shown in the far right lane (No virus). This experiment was repeated twice with comparable results.