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H H Farman Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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K L Gustafsson Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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P Henning Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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L Grahnemo Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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V Lionikaite Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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S Movérare-Skrtic Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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J Wu Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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H Ryberg Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden
Clinical Chemistry, Sahlgrenska University Hospital, Gothenburg, Sweden

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A Koskela Unit of Cancer Research and Translational Medicine, MRC Oulu and Department of Anatomy and Cell Biology, University of Oulu, Oulu, Finland

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J Tuukkanen Unit of Cancer Research and Translational Medicine, MRC Oulu and Department of Anatomy and Cell Biology, University of Oulu, Oulu, Finland

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E R Levin Division of Endocrinology, Departments of Medicine and Biochemistry, University of California, Irvine, California, USA
The Long Beach VA Medical Center, Long Beach, California, USA

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C Ohlsson Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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M K Lagerquist Centre for Bone and Arthritis Research at Institute of Medicine, Sahlgrenska Academy, University of Gothenburg, Gothenburg, Sweden

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The importance of estrogen receptor α (ERα) for the regulation of bone mass in males is well established. ERα mediates estrogenic effects both via nuclear and membrane-initiated ERα (mERα) signaling. The role of mERα signaling for the effects of estrogen on bone in male mice is unknown. To investigate the role of mERα signaling, we have used mice (Nuclear-Only-ER; NOER) with a point mutation (C451A), which results in inhibited trafficking of ERα to the plasma membrane. Gonadal-intact male NOER mice had a significantly decreased total body areal bone mineral density (aBMD) compared to WT littermates at 3, 6 and 9 months of age as measured by dual-energy X-ray absorptiometry (DEXA). High-resolution microcomputed tomography (µCT) analysis of tibia in 3-month-old males demonstrated a decrease in cortical and trabecular thickness in NOER mice compared to WT littermates. As expected, estradiol (E2) treatment of orchidectomized (ORX) WT mice increased total body aBMD, trabecular BV/TV and cortical thickness in tibia compared to placebo treatment. E2 treatment increased these skeletal parameters also in ORX NOER mice. However, the estrogenic responses were significantly decreased in ORX NOER mice compared with ORX WT mice. In conclusion, mERα is essential for normal estrogen signaling in both trabecular and cortical bone in male mice. Increased knowledge of estrogen signaling mechanisms in the regulation of the male skeleton may aid in the development of new treatment options for male osteoporosis.

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Melyssa R Bratton Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of
Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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James W Antoon Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Bich N Duong Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Daniel E Frigo Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Syreeta Tilghman Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of
Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Bridgette M Collins-Burow Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Steven Elliott Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Yan Tang Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Lilia I Melnik Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Ling Lai Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Jawed Alam Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Barbara S Beckman Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Steven M Hill Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Brian G Rowan Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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John A McLachlan Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Matthew E Burow Section of Hematology and Medical Oncology, Structural and Cellular Biology, Department of Medical Genetics, Center for Nuclear Receptors and Cell Signaling, Department of Medicine, Tulane University, 1430 Tulane Avenue, SL-78, New Orleans, Louisiana 70112, USA Departments of

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Introduction Various agents regulate estrogen receptor α (ERα) activity in addition to 17β-estradiol (E 2 ), including peptide growth factors (PGFs) such as epidermal growth factor (EGF) and insulin-like growth factor 1 (IGF1; Ignar-Trowbridge et

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Beverly A S Reyes Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Department of Neurosurgery, Farber Institute for Neurosciences, Thomas Jefferson University, 900 Walnut Street, Suite 400, Philadelphia, Pennsylvania 19107, USA
Department of Biology, Washington and Lee University, Lexington, Virginia 24450-03, USA

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Hiroko Tsukamura Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Department of Neurosurgery, Farber Institute for Neurosciences, Thomas Jefferson University, 900 Walnut Street, Suite 400, Philadelphia, Pennsylvania 19107, USA
Department of Biology, Washington and Lee University, Lexington, Virginia 24450-03, USA

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Helen I’Anson Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Department of Neurosurgery, Farber Institute for Neurosciences, Thomas Jefferson University, 900 Walnut Street, Suite 400, Philadelphia, Pennsylvania 19107, USA
Department of Biology, Washington and Lee University, Lexington, Virginia 24450-03, USA

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Maria Amelita C Estacio Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Department of Neurosurgery, Farber Institute for Neurosciences, Thomas Jefferson University, 900 Walnut Street, Suite 400, Philadelphia, Pennsylvania 19107, USA
Department of Biology, Washington and Lee University, Lexington, Virginia 24450-03, USA

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Kanjun Hirunagi Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Department of Neurosurgery, Farber Institute for Neurosciences, Thomas Jefferson University, 900 Walnut Street, Suite 400, Philadelphia, Pennsylvania 19107, USA
Department of Biology, Washington and Lee University, Lexington, Virginia 24450-03, USA

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Kei-Ichiro Maeda Graduate School of Bioagricultural Sciences, Nagoya University, Nagoya 464-8601, Japan
Department of Neurosurgery, Farber Institute for Neurosciences, Thomas Jefferson University, 900 Walnut Street, Suite 400, Philadelphia, Pennsylvania 19107, USA
Department of Biology, Washington and Lee University, Lexington, Virginia 24450-03, USA

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suppression of LH secretion. Table 1 Number of estrogen receptor α (ERα), dopamine-β-hydroxylase (DBH), and dual-labeled cells in the nucleus of the solitary tract of 48-h fasted and unfasted ovariectomized rats ( n =4) ER α DBH ER α+ DBH Double

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Liyuan Tian Molecular Biology, Endocrinology, Departments of

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Zhiqiang Wu Molecular Biology, Endocrinology, Departments of

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Yali Zhao Molecular Biology, Endocrinology, Departments of

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Yuanguang Meng Molecular Biology, Endocrinology, Departments of

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Yiling Si Molecular Biology, Endocrinology, Departments of

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Xiaobing Fu Molecular Biology, Endocrinology, Departments of

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Yiming Mu Molecular Biology, Endocrinology, Departments of

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Weidong Han Molecular Biology, Endocrinology, Departments of

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XJ Yu L Song HJ Li M Lu JM Zhao YL 2003 Up-regulation of LRP16 mRNA by 17β-estradiol through activation of estrogen receptor α (ERα), but not estrogen receptor β (ERβ), and promotes human breast cancer MCF-7 cell proliferation: a

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L Lundholm Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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G Bryzgalova
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H Gao Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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N Portwood
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S Fält Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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K D Berndt Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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A Dicker Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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D Galuska Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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J R Zierath Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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J-Å Gustafsson Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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S Efendic
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K Dahlman-Wright Department of Molecular Medicine and Surgery, Department of Biosciences and Nutrition, Department of Medicine, Department of Molecular Medicine and Surgery, Karolinska Institute, Karolinska University Hospital, SE-171 76 Stockholm, Sweden

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A Khan
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Medicine 13 89 – 94 . Gao H Falt S Sandelin A Gustafsson JA Dahlman-Wright K 2008 Genome-wide identification of estrogen receptor α-binding sites in mouse liver . Molecular Endocrinology 22 10 – 22 . Geisler JG Zawalich W Zawalich

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Jonathan Lindzey Receptor Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, MD B3-02, PO Box 12233, Research Triangle Park, North Carolina 27709, USA
Department of Biological Sciences, Lock Haven University, Lock Haven, Pennsylvania 17745, USA

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Friederike L Jayes Receptor Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, MD B3-02, PO Box 12233, Research Triangle Park, North Carolina 27709, USA
Department of Biological Sciences, Lock Haven University, Lock Haven, Pennsylvania 17745, USA

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Mariana M Yates Receptor Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, MD B3-02, PO Box 12233, Research Triangle Park, North Carolina 27709, USA
Department of Biological Sciences, Lock Haven University, Lock Haven, Pennsylvania 17745, USA

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John F Couse Receptor Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, MD B3-02, PO Box 12233, Research Triangle Park, North Carolina 27709, USA
Department of Biological Sciences, Lock Haven University, Lock Haven, Pennsylvania 17745, USA

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Kenneth S Korach Receptor Biology Section, Laboratory of Reproductive and Developmental Toxicology, National Institute of Environmental Health Sciences, National Institutes of Health, MD B3-02, PO Box 12233, Research Triangle Park, North Carolina 27709, USA
Department of Biological Sciences, Lock Haven University, Lock Haven, Pennsylvania 17745, USA

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distribution and quantitative analysis of estrogen receptor-α (ERα) and estrogen receptor-β (ERβ) messenger ribonucleic acid in the wild-type and ERα-knockout mouse. Endocrinology 138 4613 –4621. Couse JF , Bunch DO, Lindzey J

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J Varayoud Laboratorio de Endocrinología y Tumores Hormonodependientes, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, C C 242, Santa Fe, Argentina

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J G Ramos Laboratorio de Endocrinología y Tumores Hormonodependientes, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, C C 242, Santa Fe, Argentina

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L Monje Laboratorio de Endocrinología y Tumores Hormonodependientes, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, C C 242, Santa Fe, Argentina

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V Bosquiazzo Laboratorio de Endocrinología y Tumores Hormonodependientes, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, C C 242, Santa Fe, Argentina

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M Muñoz-de-Toro Laboratorio de Endocrinología y Tumores Hormonodependientes, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, C C 242, Santa Fe, Argentina

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E H Luque Laboratorio de Endocrinología y Tumores Hormonodependientes, School of Biochemistry and Biological Sciences, Universidad Nacional del Litoral, C C 242, Santa Fe, Argentina

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-Buck V & Gannon F 1998 Differentially expressed messenger RNA isoforms of the human estrogen receptor-α gene are generated by alternative splicing and promoter usage. Molecular Endocrinology 12 1939 –1954. Friend KE

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C D Simmons
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J M P Pabona
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Z Zeng
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M C Velarde
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D Gaddy
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F A Simmen
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R C M Simmen Department of Physiology and Biophysics, Arkansas Children's Nutrition Center, University of Arkansas for Medical Sciences, 4301 W Markham Street, Little Rock, Arkansas 72202, USA

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Introduction The steroid hormone 17β-estradiol (E 2 ) regulates numerous physiological and developmental processes in target tissues by binding to its cognate receptors estrogen receptor-α (ESR1) and -β (ESR2) in the presence of numerous co

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Mirja Rotinen Department of Health Sciences, Universidad Pública de Navarra, Avda. Barañain, 31008 Pamplona, Spain

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Jon Celay Department of Health Sciences, Universidad Pública de Navarra, Avda. Barañain, 31008 Pamplona, Spain

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Marta M Alonso Department of Health Sciences, Universidad Pública de Navarra, Avda. Barañain, 31008 Pamplona, Spain

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Aranzazu Arrazola Department of Health Sciences, Universidad Pública de Navarra, Avda. Barañain, 31008 Pamplona, Spain

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Ignacio Encio Department of Health Sciences, Universidad Pública de Navarra, Avda. Barañain, 31008 Pamplona, Spain

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Joaquin Villar Department of Health Sciences, Universidad Pública de Navarra, Avda. Barañain, 31008 Pamplona, Spain

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et al . 2003 , Khalyfa et al . 2003 , Bourdeau et al . 2004 ). Estrogens modulate transcription in their target tissues through estrogen receptor α (ERα) or ERβ receptors using a number of signaling pathways. The ‘classical’ pathway involves

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Rhone A Mendoza Department of Biomedical Sciences, Center of Excellence in Cancer Research, Texas Tech University Health Sciences Center, Paul L Foster School of Medicine, 5001 El Paso Drive, El Paso, Texas 79905, USA

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Marlene I Enriquez Department of Biomedical Sciences, Center of Excellence in Cancer Research, Texas Tech University Health Sciences Center, Paul L Foster School of Medicine, 5001 El Paso Drive, El Paso, Texas 79905, USA

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Sylvia M Mejia Department of Biomedical Sciences, Center of Excellence in Cancer Research, Texas Tech University Health Sciences Center, Paul L Foster School of Medicine, 5001 El Paso Drive, El Paso, Texas 79905, USA

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Emily E Moody Department of Biomedical Sciences, Center of Excellence in Cancer Research, Texas Tech University Health Sciences Center, Paul L Foster School of Medicine, 5001 El Paso Drive, El Paso, Texas 79905, USA

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Gudmundur Thordarson Department of Biomedical Sciences, Center of Excellence in Cancer Research, Texas Tech University Health Sciences Center, Paul L Foster School of Medicine, 5001 El Paso Drive, El Paso, Texas 79905, USA

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blot analysis showing expression of the estrogen receptor-α (ERα), the progesterone receptor (PR), and the ERβ in cloned MCF-7 cells stably transfected with a vector producing a small interfering RNA (siRNA) to the insulin-like growth factor-I receptor

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