Xenbase Image ID: 125041

Figure 6. Nup188–Nup93 depletion causes a faster delivery of integral membrane proteins to the INM. (A) Nuclei were assembled in mock- or Nup188–Nup93-depleted Xenopus egg extracts and analyzed by Western blotting. Positions of the uncleaved (EGFP-BC08) and the cleaved (EGFP) reporter are indicated. (B) Quantification from three independent experiments performed as in A. Squares are from Nup188–Nup93-depleted samples (green without and red with the addition of in vitro translated Nup188–Nup93 [addback]), and circles are from mock-treated samples. Lines mark the mean of three independent experiments. (C) The addition of recombinant Nup188–Nup93 rescues the Nup188–Nup93 depletion phenotype. Nuclei were assembled for 120 min in mock- or Nup188–Nup93-depleted extracts without or with the addition of in vitro translated Nup188–Nup93 (addback), fixed with 4% PFA and 0.5% glutaraldehyde, and analyzed for chromatin and membrane staining (blue, DAPI; red, DiIC18). (D) In vitro translated Nup188–Nup93 is incorporated into in vitro assembled nuclei. Nuclei were assembled for 1 h to ensure approximately equal NPC numbers per nucleus, isolated, and analyzed by Western blotting. In vitro translated Nup188 and Nup93 could be also detected using an anti-His6 antibody (third and fourth row). The nucleoporin p62 was detected with mAb414, and the chromatin-binding protein RCC1 serves as a control for equal loading of nuclei. Bar, 20 µm. Image published in: Theerthagiri G et al. (2010) © 2010 Theerthagiri et al. This image is reproduced with permission of the journal and the copyright holder. This is an open-access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike license Larger Image Printer Friendly View

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