Figure 8

Nucant pseudopeptides are internalized into cells and accumulate in the cytoplasm without translocation to the nucleus. A/B. HuT 78 cells in polylysine-coated eight-well glass slides were incubated (37°C) with 15 μM of biotinylated N3 or N7 for 5 hours before PFA/Triton fixation and processing for immunofluorescence microscopy using rabbit anti-biotin and FITC conjugated goat anti-rabbit IgG. C. HuT 78 cells in polylysine-coated slides were incubated (37°C) with 15 μM of biotinylated N7 for 5 hours before methanol/acetate fixation and processing for immunofluorescence microscopy as in A/B (Methods). The nuclei stained with DAPI are shown on the right of each panel. Treatment of HuT 78 cells at 15 μM of biotinylated N3 or N7 leads to 25 and 40% cell death after 24 of culture, respectively.