Pom121 interacts with Nup155 and the Nup107–160 complex in vitro. Bead-bound GST-Pom121215–557 or GST alone was incubated with (+) or without (−) rat liver NE extracts. Interacting proteins were eluted with SDS-sample buffer. Polypeptides were resolved by SDS-PAGE and visualized by silver staining (SS; A) or analyzed by Western blotting (WB; B) using antibodies directed against the indicated proteins. mAb414 was used to detect Nup62, Nup153, and Nup214. Approximately 5% of the NE extract loaded on each column was resolved in the lane marked Load. Prominent bands were analyzed by mass spectrometry (see Fig. S3) and the predominant species identified in the 90–160-kD range are indicated in the shaded box. Molecular mass markers are indicated in kilodaltons.
