Methods in Cell Biology Methods in Cilia & Flagella Volume 127 Series Editors Leslie Wilson DepartmentofMolecular,CellularandDevelopmentalBiology UniversityofCalifornia SantaBarbara,California Phong Tran University of Pennsylvania Philadelphia, USA & Institut Curie, Paris, France Methods in Cell Biology Methods in Cilia & Flagella Volume 127 Edited by Renata Basto InstitutCurie, France Wallace F. Marshall Department of Biochemistry & Biophysics, UCSF Mission Bay, USA AMSTERDAM(cid:129)BOSTON(cid:129)HEIDELBERG(cid:129)LONDON NEWYORK(cid:129)OXFORD(cid:129)PARIS(cid:129)SANDIEGO SANFRANCISCO(cid:129)SINGAPORE(cid:129)SYDNEY(cid:129)TOKYO AcademicPressisanimprintofElsevier AcademicPressisanimprintofElsevier 125,LondonWall,EC2Y5AS 525BStreet,Suite1800,SanDiego,CA92101-4495,USA 225WymanStreet,Waltham,MA02451,USA TheBoulevard,LangfordLane,Kidlington,OxfordOX51GB,UK Firstedition2015 Copyright©2015ElsevierInc.AllRightsReserved. Nopartofthispublicationmaybereproducedortransmittedinanyformorbyanymeans, electronicormechanical,includingphotocopying,recording,oranyinformationstorage andretrievalsystem,withoutpermissioninwritingfromthepublisher.Detailsonhowto seekpermission,furtherinformationaboutthePublisher’spermissionspoliciesandour arrangementswithorganizationssuchastheCopyrightClearanceCenterandthe CopyrightLicensingAgency,canbefoundatourwebsite:www.elsevier.com/permissions. Thisbookandtheindividualcontributionscontainedinitareprotectedundercopyrightby thePublisher(otherthanasmaybenotedherein). Notices Knowledgeandbestpracticeinthisfieldareconstantlychanging.Asnewresearchand experiencebroadenourunderstanding,changesinresearchmethods,professional practices,ormedicaltreatmentmaybecomenecessary. Practitionersandresearchersmustalwaysrelyontheirownexperienceandknowledgein evaluatingandusinganyinformation,methods,compounds,orexperimentsdescribed herein.Inusingsuchinformationormethodstheyshouldbemindfuloftheirownsafety andthesafetyofothers,includingpartiesforwhomtheyhaveaprofessionalresponsibility. Tothefullestextentofthelaw,neitherthePublishernortheauthors,contributors,or editors,assumeanyliabilityforanyinjuryand/ordamagetopersonsorpropertyasa matterofproductsliability,negligenceorotherwise,orfromanyuseoroperationofany methods,products,instructions,orideascontainedinthematerialherein. ISBN:978-0-12-802451-5 ISSN:0091-679X ForinformationonallAcademicPresspublications visitourwebsiteathttp://store.elsevier.com/ Contributors Zachary D.Abbott MountDesertIslandBiologicalLaboratory,SalisburyCove,ME,USA;Department ofBiology,Bates College, Lewiston, ME, USA JeffreyD. Amack Department ofCelland Developmental Biology,State University ofNew York, Upstate Medical University,Syracuse, NY,USA Manisha Anand Department ofOphthalmology,UMASS Medical School, Worcester,MA, USA Anne Aubusson-Fleury Centre deGe´ne´tiqueMole´culaire, Institute for IntegrativeBiologyof theCell (I2BC),Universite´ Paris Saclay, CEA, CNRS, Universite´ Paris Sud, Bat26 Alle´e de laterrasse, 91 198 Gif sur YvetteCedex, France JeffreyD. Axelrod Department ofPathology,StanfordUniversity School ofMedicine,Stanford, CA, USA Juliette Azimzadeh InstitutJacquesMonod,CNRSUMR7592/Universite´ ParisDiderot,ParisCedex, France HemantBadgandi Department ofCellBiology,UT SouthwesternMedical Center,Dallas, TX,USA Cyril Basquin InstitutJacquesMonod,CNRSUMR7592/Universite´ ParisDiderot,ParisCedex, France Philippe Bastin TrypanosomeCellBiologyUnit,InstitutPasteur&CNRSURA2581,Paris,France Renata Basto Cell Biology,Institut Curie,UMR 144 CNRS, Paris, France Gwendolyn M. Beacham Mount Desert Island BiologicalLaboratory, Salisbury Cove, ME, USA; HonorsCollege, University ofMaine,Orono,ME, USA Oliver E.Blacque School ofBiomolecular and BiomedicalScience, University College Dublin, Belfield,Dublin, Ireland xiii xiv Contributors Thomas C.Boothby Department ofCell Biologyand Molecular Genetics, University ofMaryland, College Park MD20742, USA Francesco Boselli Institut de Ge´ne´tique etde Biologie Mole´culaire etCellulaire, Illkirch, France; Centre Nationalde la Recherche Scientifique, Illkirch, France; InstitutNational de laSante´ etde laRecherche Me´dicale,Illkirch, France; Universite´ de Strasbourg,Illkirch, France David K. Breslow Department ofMolecular and Cellular Physiology,Stanford University School of Medicine,Stanford, CA, USA Eric R.Brooks DepartmentofMolecularBiosciences,TheUniversityofTexasatAustin,Austin, TX,UnitedStates Gilles Charvin Institut de Ge´ne´tique etde Biologie Mole´culaire etCellulaire, Illkirch, France; Centre Nationalde la Recherche Scientifique, Illkirch, France; InstitutNational de laSante´ etde laRecherche Me´dicale,Illkirch, France; Universite´ de Strasbourg,Illkirch, France Nicolas Chenouard BioImageAnalysis Unit,InstitutPasteur& CNRS URA2582,Paris, France JeanCohen Centre de Ge´ne´tique Mole´culaire, Institute for Integrative Biology ofthe Cell (I2BC), Universite´ ParisSaclay,CEA, CNRS,Universite´ ParisSud, Bat 26 Alle´e de laterrasse,91 198 Gif sur Yvette Cedex, France Elisabeth Cortier Centre de Ge´ne´tique etde Physiologie Mole´culaire etCellulaire, Universite´ Claude Bernard Lyon-1, Lyon, Villeurbanne,France NathalieDelgehyr Institut de Biologie de l’Ecole Normal Supe´rieure(IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS,UMR 8197, Paris, France Nicolas Diguet Institut Pasteur,Department of Developmental and Stem Cell Biology, Paris, France; CNRS URA2578,Paris, France Contributors xv Elyse M. Doherty DepartmentofBiology,WheatonCollege,Norton,MA,USA;MountDesertIsland BiologicalLaboratory,Salisbury Cove, ME, USA Be´ne´dicteDurand Centre deGe´ne´tiqueet de PhysiologieMole´culaire etCellulaire, Universite´ Claude Bernard Lyon-1, Lyon, Villeurbanne,France Susan K.Dutcher Department ofGenetics, Washington University, St.Louis,MO, USA Jean-LucDuteyrat Centre deGe´ne´tiqueet de PhysiologieMole´culaire etCellulaire, Universite´ Claude Bernard Lyon-1, Lyon, Villeurbanne,France Chelsea R.Ettinger Department ofBiology,Wheaton College, Norton, MA, USA Marion Faucourt Institut de Biologie de l’Ecole Normal Supe´rieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France Ce´cile Fort TrypanosomeCellBiologyUnit,InstitutPasteur&CNRSURA2581,Paris,France CraigFox Department ofPhysics, Syracuse University,Syracuse,NY,USA Jacek Gaertig Department ofCellular Biology, University ofGeorgia, Athens, GA,USA Eva Gluenz Sir William Dunn School ofPathology,University of Oxford, Oxford, UK Jacky G.Goetz Institut de Ge´ne´tique etde Biologie Mole´culaire etCellulaire,Illkirch, France; Centre National de laRechercheScientifique, Illkirch, France; Institut National de laSante´ etde la Recherche Me´dicale, Illkirch, France; Universite´ de Strasbourg, Illkirch, France Montserrat Bosch Grau Curie Institute, Orsay,France; CNRS,UMR3306, Orsay,France; INSERM, U1005,Orsay,France;INSERM,UMRS1120;Unite´ deGe´ne´tiqueetPhysiologie de l’Audition, PasteurInstitute,Paris, France xvi Contributors Masato Hoshi DivisionofNephrology,DepartmentofMedicine,WashingtonUniversitySchoolof Medicine,St Louis, MO, USA Louise Hughes Department ofBiologicaland Medical Sciences, Oxford Brookes University, Oxford, UK WielandB. Huttner MaxPlanckInstituteofMolecularCellBiologyandGenetics,Dresden,Germany Hiroaki Ishikawa Department ofBiochemistry and Biophysics, University of California, San Francisco, CA,USA Sanjay Jain DivisionofNephrology,DepartmentofMedicine,WashingtonUniversitySchoolof Medicine,St Louis, MO, USA; Department ofPathology and Immunology, WashingtonUniversity School ofMedicine, StLouis,MO, USA Carsten Janke Curie Institute,Orsay, France; CNRS, UMR3306,Orsay,France;INSERM, U1005, Orsay, France Yu-Yang Jiang Department ofCellular Biology,University ofGeorgia,Athens, GA, USA Ritsu Kamiya Department ofLife Science,Facultyof Science,Gakushuin University,Tokyo, Japan Julie Kennedy School ofBiomolecular and Biomedical Science,University College Dublin, Belfield, Dublin, Ireland HemantKhanna Department ofOphthalmology,UMASS Medical School,Worcester, MA, USA Thibault Lagache BioImageAnalysis Unit,InstitutPasteur& CNRS URA2582,Paris, France Karl Lechtreck Department ofCellular Biology,University ofGeorgia,Athens, GA, USA YinLoon Lee Department ofBiology,Stanford University,Stanford, CA,USA; Department of Genetics, Stanford University School ofMedicine, Stanford, CA, USA Contributors xvii Jean-Franc¸ois Le Garrec Institut Pasteur, Department ofDevelopmentaland Stem Cell Biology,Paris, France; CNRS URA2578,Paris, France Michel Lemullois Centre deGe´ne´tiqueMole´culaire, Institute for IntegrativeBiologyof theCell (I2BC),Universite´ Paris Saclay, CEA, CNRS, Universite´ Paris Sud, Bat26 Alle´e de laterrasse, 91 198 Gif sur YvetteCedex, France LinjingLi Department ofOphthalmology,UMASS Medical School, Worcester,MA, USA Huawen Lin Department ofGenetics, Washington University, St.Louis,MO, USA Tommaso Lucchesi Institut Pasteur, Department ofDevelopmentaland Stem Cell Biology,Paris, France;CNRSURA2578,Paris,France;SorbonneUniversite´s,UPMCUniversite´ Paris06, IFD, Paris, France Moe R. Mahjoub DivisionofNephrology,DepartmentofMedicine,WashingtonUniversitySchoolof Medicine, StLouis,MO, USA; Department ofCell Biology and Physiology, WashingtonUniversity School of Medicine,St Louis, MO, USA Teresa Mendes Maia Cell Biology,Institut Curie,UMR 144 CNRS, Paris, France M. Lisa Manning Department ofPhysics, Syracuse University,Syracuse,NY,USA; Syracuse Biomaterials Institute,Syracuse University, Syracuse, NY,USA WallaceF. Marshall Department ofBiochemistry and Biophysics,University ofCalifornia, San Francisco, CA, USA Sigole`neM. Meilhac Institut Pasteur, Department ofDevelopmentaland Stem Cell Biology,Paris, France; CNRS URA2578,Paris, France Alice Meunier Institut de Biologie de l’Ecole Normal Supe´rieure (IBENS), Paris, France; INSERM, U1024, Paris, France; CNRS, UMR 8197, Paris, France xviii Contributors Robert L. Morris DepartmentofBiology,WheatonCollege,Norton,MA,USA;MountDesertIsland BiologicalLaboratory, Salisbury Cove, ME, USA Saikat Mukhopadhyay Department ofCell Biology,UTSouthwestern Medical Center,Dallas, TX, USA Maxence V. Nachury Department ofMolecular and Cellular Physiology,Stanford University School of Medicine,Stanford, CA, USA Jean-Christophe Olivo-Marin BioImageAnalysis Unit,InstitutPasteur& CNRS URA2582,Paris, France Anne-MarieOrfila InstitutJacquesMonod,CNRSUMR7592/Universite´ ParisDiderot,ParisCedex, France Kasturi Pal Department ofCell Biology,UTSouthwestern Medical Center,Dallas, TX, USA Judith T.M.L. Paridaen MaxPlanckInstituteofMolecularCellBiologyandGenetics,Dresden,Germany Perrine Paul-Gilloteaux Cell Biology, Institut Curie,UMR 144 CNRS, Paris, France Hans W.Pope DepartmentofBiology,WheatonCollege,Norton,MA,USA;MountDesertIsland BiologicalLaboratory, Salisbury Cove, ME, USA Kollu Nageswara Rao Department ofOphthalmology,UMASS Medical School,Worcester, MA, USA Anna A.W.M. Sanders School ofBiomolecular and Biomedical Science,University College Dublin, Belfield, Dublin, Ireland Julien Santi-Rocca TrypanosomeCellBiologyUnit,InstitutPasteur&CNRSURA2581,Paris,France Tatsushi Shintaku DepartmentofBiology,WheatonCollege,Norton,MA,USA;MountDesertIsland BiologicalLaboratory, Salisbury Cove, ME, USA Adam N. Sholi DepartmentofBiology,WheatonCollege,Norton,MA,USA;MountDesertIsland BiologicalLaboratory, Salisbury Cove, ME, USA