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Use of Ecto-Tagged Integrins to Monitor Integrin Exocytosis and Endocytosis.
Huet-Calderwood, Clotilde; Rivera-Molina, Felix; Toomre, Derek; Calderwood, David A.
Afiliación
  • Huet-Calderwood C; Departments of Pharmacology, Yale University School of Medicine, Yale University, New Haven, CT, USA.
  • Rivera-Molina F; Departments of Cell Biology, Yale University School of Medicine, Yale University, New Haven, CT, USA.
  • Toomre D; Departments of Cell Biology, Yale University School of Medicine, Yale University, New Haven, CT, USA.
  • Calderwood DA; Departments of Pharmacology, Yale University School of Medicine, Yale University, New Haven, CT, USA. david.calderwood@yale.edu.
Methods Mol Biol ; 2608: 17-38, 2023.
Article en En | MEDLINE | ID: mdl-36653699
Controlled exocytosis and endocytosis of integrin adhesion receptors is required for normal cell adhesion, migration, and signaling. In this chapter, we describe the design of functional ß1 integrins carrying extracellular fluorescent or chemically traceable tags (ecto-tag) and methods for their use to image ß1 integrin trafficking in cells. We provide approaches to generate cells in which endogenous ß1 integrins are replaced by ecto-tagged integrins containing a pH-sensitive fluorophore pHluorin or a HaloTag and describe strategies using photobleaching, selective extracellular/intracellular labeling, and chase, quenching, and blocking to reveal ß1 integrin exocytosis, endocytosis, and recycling by live total internal reflection fluorescence (TIRF) microscopy.
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Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Integrinas / Integrina beta1 Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos

Texto completo: 1 Banco de datos: MEDLINE Asunto principal: Integrinas / Integrina beta1 Idioma: En Revista: Methods Mol Biol Asunto de la revista: BIOLOGIA MOLECULAR Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos