FIG. 7.

Inhibitors of the NLS pathway do not inhibit GST-RanBP1(1-161) import. (A) BHK-21 cells were permeabilized with 0.008% digitonin and overlaid with either 1.5 μM GST-GFP-NLS (a to e) or 3 μM GST-RanBP1(1-161) (f to j) in the presence of an energy-regenerating system, reticulocyte lysate (Retic), 100 U of apyrase/ml, 2 μM importin (imp) β(45-462)), 100 mg of RanG19V/ml, or 200 mg of WGA/ml. Cells were incubated at 23°C for 30 min and then fixed. GST-GFP-NLS and GST-RanBP1(1-161) were detected by epifluorescence and by indirect immunofluorescence with anti-GST MAb, respectively. (B) The average total fluorescence per cell of GST-RanBP1(1-161) was quantitated using Openlab software. Error bars represent the standard error of the mean (n > 20 cells per treatment).