FIG 2.

(A) Construction of different NPC truncations in the pEGFP-C1 vector. Numbers represent corresponding amino acid residues on the protein. Constructs are not drawn to scale. (B and C) Constructs in panel A were transfected into 293T cells preseeded on glass slides. At the indicated time points, cells were harvested and fixed for visualization of the GFP signal using a Nikon E80i microscope. A deletion of 12 amino acids from the C terminus of NPC did not affect localization, but deletions of 39, 51, and 60 amino acids impaired nuclear targeting. (C) C-terminal domain (CTD) alone was also able to cause nuclear import. Nuclei were stained with DAPI. Subcellular localization was classified as N (exclusively nuclear), N+C (distributed), or C≫N (mainly cytoplasmic). Percentage values reported represent average percentages of nuclear signal for 20 cells counted.