Figure 7.

Immunoblot analysis of yeast subcellular fractionation. Samples of total yeast cells (lane 1), crude cytosol (lane 2), crude nuclei/membranes (lane 3), and sucrose gradient fractions resolving the crude nuclei/membrane fraction (lanes 4–8) were prepared as described in MATERIALS AND METHODS, separated electrophoretically on a 13% SDS-polyacrylamide gel, and transferred to nitrocellulose. The blot was divided in thirds, and the top portion was probed with an affinity-purified antibody recognizing Nup116p, the middle portion with antibody for Kar2p, and the bottom portion with antibody for Snl1p. All three proteins enriched in the crude nuclei/membrane fraction. Loading equivalents represent the number of cell equivalents that were used as starting material to prepare each of the samples.