Supplemental Material

This article contains the following supporting material:

• Figure S1 - Thin section electron microscopy reveals perturbations in NE and cellular membranes of the kap95· GFP-nic96 nup170-GFP +pGAL-KAP95 mutant (SWY3189). Cells were grown to early log phase in 2% galactose, then shifted to 2% glucose for 6 hours before being processed for TEM as in Fig. 6. Arrowheads point to NPCs that span the NE, asterisks(*) mark areas where NPC-like structures associate with only the INM. Lines point to NE; arrows point to stretches of cytoplasmic (non-nuclear) membranes. N, nucleus; C, cytoplasm, V, vesicles; m, mitochondria; vac, vacuole: and cl, cluster of NPCs. All bars equal 1 μm.
• Figure S2 - Ran localization is not perturbed in the kap95-E126K GFP-nic96 nup170-GFP mutant. Mutant (SWY3561) and parental (SWY2089) cells were grown to early log phase at 23°C, shifted to growth at 34°C for 6 hours, and then processed for indirect immunfluorescence microscopy. A monoclonal anti-Ran antibody was used to detect Ran localization.