Supplemental Figures

This article contains the following supporting material:

• Supplemental Figure S1 - Mlp1p and Mlp2p physically interact with Nup145p and members of the Nup53p-containing complex.

  Mlp1-pA and Mlp2-pA associated protein complexes were immunopurified from yeast whole cell lysates, resolved by SDS-PAGE and stained with Coomassie blue (left). The vertical black bars indicate gel sections containing prominent protein bands of MW >45 kDa that were excised and used to identify peptides by LCQ-MS/MS following in-gel trypsin digestion. The identified peptides in each of the gel slices were compiled and this list was then thresholded to contain only the Mlp proteins and nups for which 9 or greater peptides were identified (right). This revealed an enrichment of several NPC core proteins in these immunopurification eluates.

• Supplemental Figure S2 - The kinetochore association of the C-terminal region of Mad1p is dependent on its NLS.

  Plasmids expressing 475-749-GFP (p475-749-GFP), 475-749-ΔNLS-GFP (p475-749-ΔNLS-GFP), 318-749-GFP (p318-749-GFP), or 318-749-ΔNLS-GFP (p318-749-ΔNLS-GFP) were introduced into a mad1Δ strain expressing Mtw1-RFP (Y3031). Asynchronous cultures or cells arrested with 12.5 μg/ml nocodazole at 30°C for 1.5 hrs (+Noc) were examined using a confocal microscope to visual the GFP and RFP fusion proteins. Merged images are shown on the right.