The LINC Complex, Softcover reprint of the original 1st ed. 2018
Methods and Protocols
Methods in Molecular Biology Series, Vol. 1840

This volume describes research methodologies and approaches used to study the Linker of Nucleoskeleton and Cytoskeleton (LINC) complex and its cellular functions. Chapters detail structural and biochemical analysis of LINC complexes, mechanical aspects of the LINC complex, analysis of the LINC complex in model systems and development, and LINC complex in mammalian tissue, organs, and disease. Written in the highly successful Methods in Molecular Biology series format, chapters include introductions to their respective topics, lists of the necessary materials and reagents, step-by-step, readily reproducible laboratory protocols, and tips on troubleshooting and avoiding known pitfalls.

Authoritative and cutting-edge, The LINC Complex: Methods and Protocols aims to ensure successful results in the further study of this vital field.

Part 1 - Structural and Biochemical Analysis of LINC complexes

1. EM Tomography of LINC Complexes

            Marie-Christin Spindler, Frederick Helmprobst, Christian Stigloher and Ricardo Benavente

2. Recombinant Purification of the Periplasmic Portion of the LINC Complex

            Victor E. Cruz and Thomas U. Schwartz

3. Analysis of High Molecular Weight Isoforms of Nesprin-1 and Nesprin-2 with Vertical Agarose Gel Electrophoresis

            Chloe Potter and Didier Hodzic

4. Analysis of Nesprin-2 Interaction with its Binding Partners and Actin

            Susumu Antoku and Gregg G. Gundersen

5. Interactions of Nesprin-4-containing LINC Complexes in Outer Hair Cells Explored by BioID

            Brian Burke

Part 2 – Mechanical Aspects of the LINC Complex

6. Using Nesprin Tension Sensors to Measure Force on the LINC Complex

            Paul T. Arsenovic and Daniel E. Conway

7. Analyzing Mechanotransduction through the LINC Complex in Isolated Nuclei

            Nejma Belaadi, Angelique Millon-Frémillon, Julien Aureille, and Christophe Guilluy

8. Direct Force Probe for Nuclear Mechanics

9. Centrifugal Displacement of Nuclei in Adherent Cells to Study LINC Complex Dependent Mechanisms of Homeostatic Nuclear Positioning

            Ruijun Zhu and Gregg G. Gundersen

10. Assembly and Use of a Microfluidic Device to Study Cell Migration in Confined Environments

            Jeremy Keys, Aaron Windsor, and Jan Lammerding

Part 3 - Analysis of the LINC Complex in Model Systems and Development

11. Investigating LINC Complex Protein Homo-oligomerization in the Nuclear Envelopes of Living Cells Using Fluorescence Fluctuation Spectroscopy

Jared Hennen, Isaac Angert, Kwang-Ho Hur, G.W. Gant Luxton, and Joachim D. Mueller

12. Functional Analysis of the Yeast LINC Complex Using Fluctuation Spectroscopy and Super Resolution Imaging

Jay R. Unruh, Brian D. Slaughter, and Sue L. Jaspersen

13. Genetic Analysis of Nuclear Migration and Anchorage to Study LINC Complexes During Development of Caenorhabditis elegans

            Heidi N. Fridolfsson, Leslie A. Herrera, James N. Brandt, Natalie E. Cain, Greg J. Hermann, and Daniel A. Starr

14. High-resolution Imaging Methods to Analyze LINC Complex Function During Drosophila Muscle Development

15. Computational Methods for Studying the Plant Nucleus

Axel Poulet, Xiao Zhou, Kentaro Tamura, Iris Meier, Christophe Tatout, Katja Graumann, and David E. Evans

            Maxime Voisin, Emmanuel Vanrobays, and Christophe Tatout

17. Immunolabeling Protocols for Studying Meiosis in Plant Mutants Defective for Nuclear Envelope Components

Javier Varas and Mónica Pradillo

Part 4 - LINC Complex in Mammalian Tissue, Organs and Disease

18. Generation and Analysis of Striated Muscle Selective LINC Complex Protein Mutant Mice

            Matthew J Stroud, Xi Fang, Jennifer Veevers, and Ju Chen

19. An in vitro System to Measure the Positioning, Stiffness and Rupture of the Nucleus in Skeletal Muscle

William Roman, Mafalda R. Pimentel, and Edgar R. Gomes

20. Functional Analysis of LINC Complexes in Skin

            Iakowos Karakesisoglou, Carmen Mroß, and Angelika A. Noegel

21. Detection of SUN1 Splicing Variants at the mRNA and Protein Levels in Cancer

            Ayaka Matsumoto, Nariaki Matsuura, and Miki Hieda

22. Next-generation Sequencing and Mutational Analysis: Implications for Genes Encoding LINC Complex Proteins

Includes cutting-edge methods and protocols

Provides step-by-step detail essential for reproducible results

Contains key notes and implementation advice from the experts