Supplemental Material

This article contains the following supporting material:

• Table 1
• Table 2
• Table 3
• Figure 1 - The Nup107-160 complex localizes to RanGTP-induced microtubule asters.
  Microtubule asters were assembled by adding RanQ69L to CSF extract along with rhodamine-labeled Tubulin. Reactions were fixed with formaldehyde, then the asters were centrifuged onto coverslips, post-fixed in methanol, and processed for immunofluorescence. The asters were probed for the presence or absence of Nup133 (green panel, first row), Nup62 (green panel, second row), or IgG (green panel, bottom row). The presence of rhodamine-labeled Tubulin (red, all rows) is also shown. The bar represents 10 microns.

• Figure 2 - The Nup107-160 complex is not required for induction of aster or microspindle assembly by Ran-GTP.
  RanQ69L was added to either mock-depleted or Nup107-160 complex-depleted Xenopus mitotic (CSF) extracts along with rhodamine-labeled Tubulin. Aliquots of reactions were examined at 60 minutes with a Zeiss Axioskop 2 microscope. Mock (left panel) and Nup107-160 complex-depleted (right panel) extracts showed equally robust aster assembly. Similarly, both extracts were capable of forming bipolar spindle-like structures by combining asters under chromatin-free conditions. The bar represents 10 microns.