Figure 3

Nuclear import mediated by recombinant HIV-1 IN protein: studies with microinjected and permeabilized mammalian cells. Solutions containing the following conjugates: (A) FITC-BSA-IN, (B) FITC-BSA-180-IN, (C) FITC-BSA-NLS-IN, (D) FITC-BSA-152-IN and (E) FITC-BSA*, were microinjected into the cytoplasm of cultured COS-7 cells. All other experimental conditions were as described in Methods. (F) Nuclear import was quantitatively estimated by an ELISA-based assay system. Digitonin-permeabilized Colo-205 cells were incubated for 1 h with Bb-IN, Bb-NLS-IN or Bb-152-IN (4 μg) in a final volume of 40 μL of transport buffer containing ATP regeneration system. The nuclear import experiments were repeated at least three times; data given in the figure represent results obtained from a single experiment. Error bars represent standard deviation which is about +/-5%. Bar 10 μm.