Figure - PMC

Skip to main content

An official website of the United States government

Here's how you know

Here's how you know

Official websites use .gov
A .gov website belongs to an official government organization in the United States.

Secure .gov websites use HTTPS
A lock ( ) or https:// means you've safely connected to the .gov website. Share sensitive information only on official, secure websites.

View full-text article in PMC

. 2010 Nov 1;191(3):505–521. doi: 10.1083/jcb.201007098

Search in PMC
Search in PubMed
View in NLM Catalog
Add to search

© 2010 Mitchell et al.

This article is distributed under the terms of an Attribution–Noncommercial–Share Alike–No Mirror Sites license for the first six months after the publication date (see http://www.rupress.org/terms). After six months it is available under a Creative Commons License (Attribution–Noncommercial–Share Alike 3.0 Unported license, as described at http://creativecommons.org/licenses/by-nc-sa/3.0/).

PMC Copyright notice

Figure 4. — Identification of Nup155-interacting proteins. (A) GST-Nup155 or GST alone were bound to glutathione–Sepharose beads and then incubated in the presence (+) or absence (−) of solubilized rat liver NE extracts. Bound proteins were eluted from beads using SDS-sample buffer. Interacting nups were detected by Western blot (WB) using antibodies directed against the indicated proteins. mAb414 was used to detect Nup62 and Nup153. Approximately 5% of the NE extract loaded on each column was resolved in the lane marked Load. (B) GST-tagged truncations of the pore-facing domain of Pom121 (amino acid residues 215–557, 558–1218, or 835–1218) were incubated with (+) or without (−) rat liver NE extracts. Interacting proteins were eluted with SDS-sample buffer. Polypeptides were resolved by SDS-PAGE and analyzed by Western blotting (WB) using a Nup155-specific polyclonal antibody. Approximately 5% of the NE extract loaded on each column was resolved in the lane marked Load. (C and D) Glutathione–Sepharose beads with attached GST-Nup155, GST-Nup53, or GST alone were incubated with (+) or without (−) purified recombinant Pom121^215–557 (C) or Ndc1^292–674 (D). Bound proteins were processed as described in A and polypeptides were detected using Coomassie blue staining (CB) or Western blot (WB) using anti-NDC1 antibodies. The lanes marked Load contain ∼50% of the total purified Pom121^215–557 (C) or NDC1^292–674 (D) loaded on the beads. To the right of each panel, the point at which the named recombinant protein migrates in the appropriate lane is indicated. Mass markers are in kilodaltons.