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Eric Y Lian Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Kingston, Ontario, Canada
Department of Pathology & Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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Sarah M Maritan Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Kingston, Ontario, Canada
Department of Pathology & Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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Jessica G Cockburn Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Kingston, Ontario, Canada
Department of Pathology & Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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Katayoon Kasaian Michael Smith Genome Sciences Centre, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada

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Mathieu J F Crupi Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Kingston, Ontario, Canada
Department of Pathology & Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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David Hurlbut Department of Pathology & Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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Steven J M Jones Michael Smith Genome Sciences Centre, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada
Department of Medical Genetics, University of British Columbia, British Columbia Cancer Research Centre, Vancouver, British Columbia, Canada

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Sam M Wiseman Department of Surgery, St Paul’s Hospital & University of British Columbia, Vancouver, British Columbia, Canada

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Lois M Mulligan Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Kingston, Ontario, Canada
Department of Pathology & Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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share the first 1063 residues but have 9 or 51 unique C-terminal amino acids, respectively ( Mulligan 2014 ). RET isoforms are generally co-expressed but have distinct molecular and functional properties. RET9 and RET51 differ in membrane localisation

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Tirtha K Das Department of Cell, Developmental and Regenerative Biology, School of Biomedical Sciences, Icahn School of Medicine, New York, New York, USA

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Ross L Cagan Department of Cell, Developmental and Regenerative Biology, School of Biomedical Sciences, Icahn School of Medicine, New York, New York, USA

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: genetic screens To model RET M918T (the RET isoform associated with MEN2B) Drosophila RET M955T was targeted to the developing eye, a well characterized epithelia in terms of cell–cell interactions and signal transduction ( Read et al . 2005 ). RET

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Maria Grazia Borrello Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Antonella Aiello Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Bernard Peissel Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Maria Grazia Rizzetti Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Piera Mondellini Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Debora Degl'Innocenti Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Veronica Catalano Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Morena Gobbo Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Paola Collini Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Italia Bongarzone Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Marco A Pierotti Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Angela Greco Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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Ettore Seregni Operative Unit Molecular Mechanisms, Pathology and Laboratory Medicine, Unit of Medical Genetics, Scientific Directorate, Nuclear Medicine, Department of Experimental Oncology

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performed on pCDNA3 eukaryotic expression vector carrying the two RET isoforms ( RET9 or RET51 ) to obtain the following mutants: RET9 -K666E, RET51 -K666E, RET9 -G691S, RET51 -G691S, RET9 -K666E–G691S, and RET51 -K666E–G691S. To assess the quality

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Giovanni Vitale Department of Clinical Sciences and Community Health (DISCCO), University of Milan, Milan, Italy
Laboratory of Endocrine and Metabolic Research, Istituto Auxologico Italiano IRCCS, Milan, Italy

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Germano Gaudenzi Department of Clinical Sciences and Community Health (DISCCO), University of Milan, Milan, Italy

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Luisa Circelli Department of Experimental Oncology, Laboratory of Molecular Biology and Viral Oncology, Istituto Nazionale per lo Studio e la Cura dei Tumori, ‘Fondazione Pascale’ – IRCCS, Naples, Italy

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Marco F Manzoni Department of Endocrinology and Internal Medicine, Endocrine Tumors Unit, San Raffaele Hospital Vita-Salute San Raffaele University, Milan, Italy

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Andrea Bassi Department of Physics, Politecnico di Milano, Milan, Italy

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Niccolò Fioritti Department of Biosciences, University of Milan, Milan, Italy

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Antongiulio Faggiano Thyroid and Parathyroid Surgery Unit, Istituto Nazionale per lo Studio e la Cura dei Tumori ‘Fondazione G. Pascale’ – IRCCS, Naples, Italy

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Annamaria Colao Department of Clinical Medicine and Surgery, Section of Endocrinology, ‘Federico II’ University of Naples, Naples, Italy

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on behalf of NIKE Group Department of Clinical Sciences and Community Health (DISCCO), University of Milan, Milan, Italy

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Ret gene ( Table 1 ). Table 1 Genetically engineered mouse models of medullary thyroid cancer carrying RET mutations. RET mutation RET isoform Promoter Background strain Tumor phenotype Reference C634R RET9 Rat

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Francesca Carlomagno
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Teresa Guida
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Suresh Anaganti
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Livia Provitera
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Svend Kjaer Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Structural Biology Laboratory, Cancer Discovery, c/o Dipartimento di Biologia e Patologia Cellulare e Molecolare, Università di Napoli Federico II, via S. Pansini 5, 80131 Naples, Italy

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Neil Q McDonald Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Structural Biology Laboratory, Cancer Discovery, c/o Dipartimento di Biologia e Patologia Cellulare e Molecolare, Università di Napoli Federico II, via S. Pansini 5, 80131 Naples, Italy

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Anderson J Ryan Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Structural Biology Laboratory, Cancer Discovery, c/o Dipartimento di Biologia e Patologia Cellulare e Molecolare, Università di Napoli Federico II, via S. Pansini 5, 80131 Naples, Italy

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Massimo Santoro
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type . Japanese Journal of Cancer Research 90 1231 – 1237 . Vidal M Wells S Ryan A Cagan R 2005 ZD6474 suppresses oncogenic RET isoforms in a Drosophila model for type 2 multiple endocrine neoplasia syndromes and papillary thyroid

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Donata Vitagliano
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Valentina De Falco
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Anna Tamburrino
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Sabrina Coluzzi
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Giancarlo Troncone Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Istituto Nazionale dei Tumori di Napoli, Division of Medical Oncology, Dipartimento di Endocrinologia e Oncologia Molecolare e Clinica, Department of Medicine and Human Oncology and Pathogenesis Program, Cancer Discovery, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, Via S. Pansini 5, 80131 Napoli, Italy

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Gennaro Chiappetta Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Istituto Nazionale dei Tumori di Napoli, Division of Medical Oncology, Dipartimento di Endocrinologia e Oncologia Molecolare e Clinica, Department of Medicine and Human Oncology and Pathogenesis Program, Cancer Discovery, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, Via S. Pansini 5, 80131 Napoli, Italy

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Fortunato Ciardiello Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Istituto Nazionale dei Tumori di Napoli, Division of Medical Oncology, Dipartimento di Endocrinologia e Oncologia Molecolare e Clinica, Department of Medicine and Human Oncology and Pathogenesis Program, Cancer Discovery, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, Via S. Pansini 5, 80131 Napoli, Italy

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Giampaolo Tortora Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Istituto Nazionale dei Tumori di Napoli, Division of Medical Oncology, Dipartimento di Endocrinologia e Oncologia Molecolare e Clinica, Department of Medicine and Human Oncology and Pathogenesis Program, Cancer Discovery, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, Via S. Pansini 5, 80131 Napoli, Italy

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James A Fagin Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Istituto Nazionale dei Tumori di Napoli, Division of Medical Oncology, Dipartimento di Endocrinologia e Oncologia Molecolare e Clinica, Department of Medicine and Human Oncology and Pathogenesis Program, Cancer Discovery, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, Via S. Pansini 5, 80131 Napoli, Italy

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Anderson J Ryan Dipartimento di Biologia e Patologia Cellulare e Molecolare, Dipartimento di Scienze Biomorfologiche e Funzionali, Istituto Nazionale dei Tumori di Napoli, Division of Medical Oncology, Dipartimento di Endocrinologia e Oncologia Molecolare e Clinica, Department of Medicine and Human Oncology and Pathogenesis Program, Cancer Discovery, Istituto di Endocrinologia ed Oncologia Sperimentale del CNR, Università Federico II, Via S. Pansini 5, 80131 Napoli, Italy

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Francesca Carlomagno
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Massimo Santoro
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clinical applications . Nature Clinical Practice. Oncology 5 521 – 530 doi:10.1038/ncponc1161 . Vidal M Wells S Ryan A Cagan R 2005 ZD6474 suppresses oncogenic RET isoforms in a Drosophila model for type 2 multiple endocrine neoplasia

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Maria Domenica Castellone Istituto di Endocrinologia ed Oncologia Sperimentale del CNR ‘G. Salvatore’, Naples, Italy

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Rosa Marina Melillo Istituto di Endocrinologia ed Oncologia Sperimentale del CNR ‘G. Salvatore’, Naples, Italy
Dipartimento di Medicina Molecolare e Biotecnologie Mediche, University of Naples ‘Federico II’, Naples, Italy

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2002 ). Three RET isoforms (RET9, RET43 and RET51) encoding for protein variants differing in the intracellular tyrosines involved in RET activation ( Tahira et al . 1990 , Lorenzo et al . 1995 , Matera et al . 2000 ) have been described. RET is

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Lois M Mulligan Division of Cancer Biology and Genetics, Cancer Research Institute, Queen’s University, Kingston, Ontario, Canada
Department of Pathology and Molecular Medicine, Queen’s University, Kingston, Ontario, Canada

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Rekab AN Lian EY Wagner SM Antonescu CN Mulligan LM 2017 Differential recruitment of E3 ubiquitin ligase complexes regulates RET isoform internalization . Journal of Cell Science 130 3282 – 3296 . ( https://doi.org/10.1242/jcs.203885 ) 28794017

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Hugo Prazeres Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal
Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal
Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Joana P Couto Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Fernando Rodrigues Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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João Vinagre Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Joana Torres Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Vitor Trovisco Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Teresa C Martins Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Manuel Sobrinho-Simões Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal
Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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Paula Soares Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal
Institute of Molecular Pathology and Immunology of the University of Porto (IPATIMUP) – Cancer Biology, Molecular Pathology Service, Endocrinology Service of the Portuguese Institute of Oncology of Coimbra FG, Department of Pathology, Rua Dr Roberto Frias, s/n, 4200-465 Porto, Portugal

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screening was performed in DNA obtained from peripheral blood leucocytes, by PCR amplification and direct Sanger sequencing of exons 8, 10, 11, and 13–16. Site-directed mutagenesis A pRcCMV vector expressing RET isoform 51 (i51) was mutated to generate the

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Lucieli Ceolin Thyroid Unit, Endocrine Division, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brasil

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Marta Amaro da Silveira Duval Thyroid Unit, Endocrine Division, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brasil

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Antônio Felippe Benini Thyroid Unit, Endocrine Division, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brasil

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Carla Vaz Ferreira Thyroid Unit, Endocrine Division, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brasil

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Ana Luiza Maia Thyroid Unit, Endocrine Division, Hospital de Clínicas de Porto Alegre, Universidade Federal do Rio Grande do Sul, Porto Alegre, Rio Grande do Sul, Brasil

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three RET isoforms, but the tyrosine residue 1096 is present only in the long (RET 51) isoform. RET is a receptor tyrosine kinase essential for the normal development and maturation of different tissues. Under normal conditions, RET is activated by a

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