Part I: The Nuclear Pore Complex and Nucleoporins

1. Affinity Isolation of Endogenous Saccharomyces cerevisiae Nuclear Pore Complexes

Ilona Nudelman, Javier Fernandez-Martinez, and Michael P. Rout

Nikola Kellner and Ed Hurt

3. Nuclear Pore Complex Assembly using Xenopus Egg Extract

Guillaume Holzer and Wolfram Antonin

Part II: Nucleo-cytoplasmic Passage

4. Analysis of Nuclear Pore Complex Permeability in Mammalian Cells and Isolated Nuclei Using Fluorescent Dextrans

5. Hormone-inducible Transport Reporter Assay to Study Nuclear Import Defects in Neurodegenerative Diseases

Saskia Hutten and Dorothee Dormann

6. Subcellular Fractionation Suitable for Studies of RNA and Protein Trafficking

Biljana Culjkovic-Kraljacic and Katherine L.B. Borden

7. Localising Total mRNA in Plant Cells

Geraint Parry

8. Using Single Molecule RNA FISH to Determine Nuclear Export and Transcription Phenotypes in Drosophila Tissues

Jennifer R. Aleman, Shawn C. Little, and Maya Capelson

Part III: Functional Analysis of Nucleoporins

9. Analysis of Nucleoporin Function using Inducible Degron Techniques

Vasilisa Aksenova, Alexei Arnaoutov, and Mary Dasso

10. Monitoring of Chromatin Organization at the Nuclear Pore Complex, Inner Nuclear Membrane and Nuclear Interior in Live Cells by Fluorescence Ratiometric Imaging of Chromatin (FRIC)

Frida Niss, Cecilia Bergqvist, Anna-Lena Ström, and Einar Hallberg

11. Analysis of Nuclear Pore Complexes in Caenorhabditis elegans by Live Imaging and Functional Genomic

Patricia de la Cruz Ruiz, Raquel Romero-Bueno, and Peter Askjaer

12. Protein Retargeting in Aspergillus nidulans to Study the Function of Nuclear Pore Complex Proteins

Subbulakshmi Suresh and Stephen A. Osmani

Part IV: Protein-Protein Interactions

13. Split-GFP Complementation to Study the Nuclear Membrane Proteome using Microscopy. Shary N. Shelton, Sarah E. Smith, and Sue L. Jaspersen

14. Bimolecular Fluorescence Complementation: Quantitative Analysis of In Cell Interaction of Nuclear Transporter Importin a with Cargo Proteins

Alexander Lee, Marie A. Bogoyevitch and David A. Jans

15. Validation of Nuclear Pore Complex Protein-Protein Interactions by Transient Expression in Plants

Fumika Ikeda and Kentaro Tamura

16. Binding affinity measurement of Nuclear Export Signal peptides to their exporter CRM1. Ho Yee Joyce Fung, and Yuh Min Chook

Part V: Post Translational Modifications

17. Analysis of Ubiquitylation and SUMOylation of Yeast Nuclear Pore Complex Proteins

Catherine Dargemont

18. Purification of Cdk-CyclinB-Kinase-Targeted Phosphopeptides from Nuclear Envelope

Justin D. Blethrow, Amanda L. DiGuilio, and Joseph S. Glavy

19. Crystallization of Nuclear Export Signals or Small Molecule Inhibitors Bound to Nuclear Exporter CRM1