Figure 6

HIV-1 Rev dominant nuclear import pathways are cell specific. Nuclear import of GST-YFP-Rev, GST-YFP-M9 and GST-SV40T NLS-GFP were examined using in vitro nuclear import assays. Digitonin permeabilized HeLa cells were incubated with 10 μl of reaction mixtures containing 1 μg of an import substrate, ATP regeneration system, and 293T/HeLa/U937/Jurkat cytosolic extracts. Recombinant 6×His-HIC at 0.5, 1 or 2 μg was added. Rev nuclear signal intensities were analyzed by ImageJ for a minimum of 100 cells and illustrated by box plots. Statistical significance analysis was performed with a two-tailed unpaired Student's t test *, P < 0.05; **, P < 0.01; ***, P < 0.001