Figure 7.

Import assays were accomplished on digitonin-permeabilized HeLa cells. Reactions consisted of GGNLS (0.2 mg/ml; top) or RCC1zz (0.7 μM; bottom) as import substrates, an energy regenerating system, and a reticulocyte lysate. Where indicated, 0.2 mg/ml WGA was added. All samples that used GGNLS as an import substrate were permeabilized with methanol and stained with a Texas red–conjugated rabbit IgG (TR-IgG; top, right panels). When RCC1zz was used as the import substrate, half of the samples were permeabilized with methanol (+MeOH) while half were not (−MeOH). RCC1zz was then visualized in all samples using a Texas red–conjugated rabbit IgG. When Triton-permeabilized cells were used (bottom, left panels), the permeabilization with methanol was omitted. All nuclei were visualized with DAPI. Bar, 10 μm.