FIGURE 5.

AES interacts in vivo with NUP98 but not with other FG repeat nucleoporins. A, full-length NUP153, NUP153 deletion mutants, NUP62, and NUP62 deletion mutants. Vertical lines represent FG repeats. B, GST or GST-AES immobilized on beads were incubated with in vitro translated NUP153, NUP62, or their deletion mutants. Bound and unbound fractions were resolved by SDS-PAGE and subjected to autoradiography. C, co-immunoprecipitation of NUP153, NUP62, and NUP98 with AES in K562 cells. Whole cell lysates were subjected to immunoprecipitation with either rabbit IgG or anti-AES antibody. Both input and immunoprecipitates (IP) were probed for NUP153 or NUP62 using the mAb414 antibody or NUP98 using anti-NUP98 antibody. AES was detected with anti-AES antibody.