title: Imaging CFTR protein localization in cultured cells and tissues.
abstract: CFTR functions as a chloride channel at the apical membrane of airway, gastrointestinal, and other epithelial cells. Immunofluorescence microscopy is commonly used to assess the subcellular localization and relative abundance of CFTR. Visualization of heterologously overexpressed CFTR is typically unproblematic and straightforward, whereas detection of small quantities of endogenous CFTR in tissues can be challenging and requires highly specific antibodies and optimized staining protocols. CFTR tagged by green fluorescent protein can be employed to study trafficking in live cells. Tagging of CFTR with an extracellular epitope permits detection exclusively at the cell surface and subsequent chasing allows visualization of endocytic trafficking.
authors.affiliation: Cystic Fibrosis/Pulmonary Research and Treatment Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, USA. silkre@med.unc.edu
authors.forename: Silvia M
authors.initials: SM
authors.lastname: Kreda
authors.forename: Martina
authors.initials: M
authors.lastname: Gentzsch
mesh_terms: D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism*; D018929:Cell Culture Techniques; D002460:Cell Line; D002462:Cell Membrane / Q000378:metabolism; D006224:Cricetinae; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000503:physiopathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000276:immunology / Q000378:metabolism*; D004847:Epithelial Cells / Q000378:metabolism / Q000648:ultrastructure; D005404:Fixatives / Q000737:chemistry; D005557:Formaldehyde / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000032:analysis* / Q000096:biosynthesis; D006801:Humans; D007150:Immunohistochemistry / Q000379:methods; D051379:Mice; D008856:Microscopy, Fluorescence / Q000379:methods*; D057054:Molecular Imaging / Q000379:methods*; D016612:Paraffin Embedding / Q000379:methods; D021381:Protein Transport; D016707:Tissue Fixation / Q000379:methods
chemical_list: D000906:Antibodies; D005404:Fixatives; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D049452:Green Fluorescent Proteins; D005557:Formaldehyde