The LINC Complex: Methods and Protocols

Front Matter ....Pages i-xiv
Front Matter ....Pages 1-1
 EM Tomography of Meiotic LINC Complexes  (Marie-Christin Spindler, Frederik Helmprobst, Christian Stigloher, Ricardo Benavente)....Pages 3-15
 Recombinant Purification of the Periplasmic Portion of the LINC Complex  (Victor E. Cruz, Thomas U. Schwartz)....Pages 17-23
 Analysis of High Molecular Weight Isoforms of Nesprin-1 and Nesprin-2 with Vertical Agarose Gel Electrophoresis  (Chloe Potter, Didier Hodzic)....Pages 25-33
 Analysis of Nesprin-2 Interaction with Its Binding Partners and Actin  (Susumu Antoku, Gregg G. Gundersen)....Pages 35-43
 Interactions of Nesprin-4-Containing LINC Complexes in Outer Hair Cells Explored by BioID  (Brian Burke)....Pages 45-56
Front Matter ....Pages 57-57
 Using Nesprin Tension Sensors to Measure Force on the LINC Complex  (Paul T. Arsenovic, Daniel E. Conway)....Pages 59-71
 Analyzing Mechanotransduction Through the LINC Complex in Isolated Nuclei  (Nejma Belaadi, Angélique Millon-Frémillon, Julien Aureille, Christophe Guilluy)....Pages 73-80
 Direct Force Probe for Nuclear Mechanics  (Vincent J. Tocco, Srujana Neelam, Qiao Zhang, Richard B. Dickinson, Tanmay P. Lele)....Pages 81-90
 Centrifugal Displacement of Nuclei in Adherent Cells to Study LINC Complex-Dependent Mechanisms of Homeostatic Nuclear Positioning  (Ruijun Zhu, Gregg G. Gundersen)....Pages 91-100
 Assembly and Use of a Microfluidic Device to Study Cell Migration in Confined Environments  (Jeremy Keys, Aaron Windsor, Jan Lammerding)....Pages 101-118
Front Matter ....Pages 119-119
 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, Joachim D. Mueller)....Pages 121-135
 Functional Analysis of the Yeast LINC Complex Using Fluctuation Spectroscopy and Super-Resolution Imaging  (Jay R. Unruh, Brian D. Slaughter, Sue L. Jaspersen)....Pages 137-161
 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, Daniel A. Starr)....Pages 163-180
 High-Resolution Imaging Methods to Analyze LINC Complex Function During Drosophila Muscle Development  (Alexander L. Auld, Mary Ann Collins, Torrey R. Mandigo, Eric S. Folker)....Pages 181-203
 Computational Methods for Studying the Plant Nucleus  (Axel Poulet, Xiao Zhou, Kentaro Tamura, Iris Meier, Christophe Tatout, Katja Graumann et al.)....Pages 205-219
 Investigation of Nuclear Periphery Protein Interactions in Plants Using the Membrane Yeast Two-Hybrid (MbY2H) System  (Maxime Voisin, Emmanuel Vanrobays, Christophe Tatout)....Pages 221-235
 Immunolabeling Protocols for Studying Meiosis in Plant Mutants Defective for Nuclear Envelope Components  (Javier Varas, Mónica Pradillo)....Pages 237-247
Front Matter ....Pages 249-249
 Generation and Analysis of Striated Muscle Selective LINC Complex Protein Mutant Mice  (Matthew J. Stroud, Xi Fang, Jennifer Veevers, Ju Chen)....Pages 251-281
 An In Vitro System to Measure the Positioning, Stiffness, and Rupture of the Nucleus in Skeletal Muscle  (William Roman, Mafalda R. Pimentel, Edgar R. Gomes)....Pages 283-293
 Functional Analysis of LINC Complexes in the Skin  (Iakowos Karakesisoglou, Carmen Mroß, Angelika A. Noegel)....Pages 295-306
 Detection of SUN1 Splicing Variants at the mRNA and Protein Levels in Cancer  (Ayaka Matsumoto, Nariaki Matsuura, Miki Hieda)....Pages 307-319
 Next-Generation Sequencing and Mutational Analysis: Implications for Genes Encoding LINC Complex Proteins  (Peter L. Nagy, Howard J. Worman)....Pages 321-336
Back Matter ....Pages 337-338