FIG. 6.

Cytosolic factor requirements. The import substrates GST-NLS-EGFP and sigmaA were dissolved together in complete transport solution (row 1) or in transport solution lacking the cytosolic extract (rows 2 to 5). The resulting solutions were added to digitonin-permeabilized cells that had been mock-incubated (rows 1 and 2) or incubated at room temperature with hypertonic buffer (row 3), NEM (row 4), and GMP-PNP (row 5) as indicated in Materials and Methods. After 30 min at 30°C, the cells were fixed, stained with DAPI (blue) and immunostained for sigmaA (red). The cells were visualized by fluorescence microscopy, and GST-NLS-GFP is stained in green.