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. 2005 Sep 29;24(20):3519–3531. doi: 10.1038/sj.emboj.7600825

Figure 4.

Figure 4

Immunodepletion of Nup155 inhibits NPC formation in vitro. (A) Nup155 was immunodepleted from cytosol and nuclei were assembled in vitro. Nucleoporin localization was analysed by immunofluorescence in mock-depleted (Mock) and Nup155-depleted reactions (ΔNup155), or after readdition of recombinant Nup155 (ΔNup155+Nup155). Nup155, mAb414 and overlay (Nup155 (green), mAb414 (red) and DAPI in blue) are costainings of the same nuclei. Scale bars, 5 μm. (B) Amounts of Nup155 in mock-depleted (Mock) and Nup155-depleted (ΔNup155) cytosol was compared by Western blotting. Equal volumes were loaded while varying amounts of untreated cytosol (Cyt) were loaded for comparison. A 0.6 μl portion of cytosol (25 μg total protein) corresponds to 50% of the sample volume loaded of mock- and Nup155-depleted cytosol. Tubulin serves as a loading control. (C, D) As in panel A, but co-immunodetection of Nup107 (C) or Pom121 (D) with mAb414 antigens.