Figure 7.

Nup96 stably interacts with the Nup107-160 complex in mitosis and is localized at kinetochores in mitotic HeLa cells. (A) Immunoprecipitation of interphase (I) or mitotic (M) HeLa S3 cell extracts by using affinity-purified anti-Nup133 antibodies. Equivalent amounts of total extracts (T) and depleted supernatants (S) and fivefold equivalents of the immune pellets (P) were analyzed by immunoblot by using antibodies directed against Nup133, Nup96, Nup85, Nup37, and Sec13. Similar coimmunoprecipitation efficiencies were obtained with these various nucleoporins from both interphase and mitotic extracts. Note the very efficient codepletion and coprecipitation of Nup96 together with Nup133. (B) Deconvolved images of HeLa cells double-labeled with affinity-purified anti-Nup96 and monoclonal anti-CENP-F antibodies. DNA was stained with DAPI. Note that Nup96 is clearly detected at kinetochores in prophase (cell on the left) and metaphase.