title: Proximity ligation assay to detect and localize the interactions of ERα with PI3-K and Src in breast cancer cells and tumor samples.
abstract: In situ proximity ligation assay (PLA) is a powerful method for detection, localization, and quantification of proteins, protein-protein interactions, and posttranslational modifications. Proteins detected by two specific antibodies are recognized by proximity probes conjugated with complementary oligonucleotides to allow the formation of circular DNA probes when bound in close proximity. Subsequent amplification of this DNA can then be visualized. Here, we describe the in situ PLA method for the detection of the ERα/Src/PI3K complex in breast cancer. We used two different techniques for detecting the signals: fluorescent detection for cell line analysis and bright-field revelation, which is better suited to clinical analysis of patient samples.
authors.affiliation: Cancer Research Center of Lyon, Centre Léon Bérard, UMR INSERM 1052, CNRS 5286, 28, Rue Laënnec, 69373, Lyon Cedex 08, France.
authors.forename: Coralie
authors.initials: C
authors.lastname: Poulard
authors.forename: Juliette
authors.initials: J
authors.lastname: Rambaud
authors.forename: Muriel
authors.initials: M
authors.lastname: Le Romancer
authors.forename: Laura
authors.initials: L
authors.lastname: Corbo
mesh_terms: D001940:Breast / Q000378:metabolism / Q000473:pathology; D001943:Breast Neoplasms / Q000378:metabolism* / Q000473:pathology; D047628:Estrogen Receptor alpha / Q000378:metabolism*; D005260:Female; D006801:Humans; D061986:MCF-7 Cells; D008856:Microscopy, Fluorescence / Q000379:methods; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism*; D025941:Protein Interaction Mapping / Q000379:methods*; D060066:Protein Interaction Maps; D019061:src-Family Kinases / Q000378:metabolism*
chemical_list: D047628:Estrogen Receptor alpha; D019869:Phosphatidylinositol 3-Kinases; D019061:src-Family Kinases; C506487:ESR1 protein, human