Figure 4.

The middle and C‐terminal region of Nup188 are responsible for chromosome alignment. (a) Expression of GFP‐tagged deletion mutants. Lysates were prepared from HeLa cells transfected with RNAi‐resistant GFP‐Nup188 mutants for 24 h, preceded by transfection with Nup188 siRNA for 36 h. Cell lysates were separated using SDS‐PAGE and probed using western blotting with an anti‐GFP or anti‐actin antibody. Arrowheads indicate the bands of corresponding mutants. (b) Representative images of cells expressing GFP, GFP‐Nup188‐Full and GFP‐Nup188‐delta N as well as mock‐treated cells. HeLa cells were transfected with RNAi‐resistant GFP‐Nup188 constructs for 24 h, preceded by transfection with Nup188 siRNA for 36 h. Cells were treated with MG132 for the final 2 h. DNA was stained with DAPI. Bar, 10 μm. (c) The percentage of cells with misaligned chromosomes for cells expressing each construct shown in (a). HeLa cells were treated as in (b). Error bars represent the SD.