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. 2001 Sep 17;154(6):1147–1160. doi: 10.1083/jcb.200101081

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Copyright © 2001, The Rockefeller University Press

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Figure 2. — Immunoprecipitation of HeLa cell extracts using anti-hNup107 or anti-hNup133 antibodies. (A) Immunoprecipitation of HeLa interphasic cell extract using preimmune (−) or immune (+) sera directed against the NH₂-terminal domain of hNup107 or against hNup133. Equivalent amounts of total extracts (T) and immune supernatants (S) and a fivefold equivalent of the immune pellets (P) were analyzed by immunoblot using anti-hNup133, hNup107, or the mAb414 antibody that mainly recognized p62. (B) Silver staining of immunoprecipitates from HeLa cell extracts obtained with preimmune (−) or immune (+) anti-hNup107 serum. (C and D) Same as A, using interphasic (I) or mitotic (M) extracts from HeLa cells. Note that hNup133 migrates as a more diffuse band in the mitotic extracts and that the slowly migrating forms of hNup33 are also precipitated efficiently by the anti-hNup107 antibody. (E) Interphasic (I) or mitotic (M) immune pellets obtained with affinity purified anti-hNup133 antibodies were analyzed by silver staining. In both samples, the same four prominent bands corresponding to hNup120, hNup133, hNup96, and hNup107 are present. In the immune pellet from mitotic extract, band 3 migrates as a doublet (*).