Testosterone (T), 17β-estradiol (E2), dibutylphthalate (DBP), flutamide (FLUT), and fulvestrant (ICI182,780) were purchased from Sigma-Aldrich (Sigma Aldrich, St. Louis, MO, United States) and dissolved in DMSO (Sigma Aldrich, St. Louis, MO, United States). T, E2, and DBP were diluted in RPMI 1640 red-phenol free (Sigma Aldrich, St. Louis, MO, United States), reaching final concentrations from 10−6 to 10−12 M, and mixed to create the experimental class used in this study. Instead, ICI182,780 and FLUT were diluted in RPMI 1640 red-phenol free at final concentrations of 10−5 M and 10−7 M, respectively.
Fulvestrant ici182 780
Manufactured by Merck Group
Sourced in Germany
Fulvestrant (ICI182,780) is a selective estrogen receptor down-regulator (SERD) that is used as a laboratory tool for research purposes. It functions by binding and degrading the estrogen receptor, thereby inhibiting estrogen-mediated signaling pathways.
Automatically generated - may contain errors
Lab products found in correlation
17β estradiol, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
Penicillin streptomycin p s, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Dmem f12, by Thermo Fisher Scientific (1 mentions)
Diethanolamine, by Merck Group (1 mentions)
Rhtgf β1, by R&D Systems (1 mentions)
Rhil 6, by R&D Systems (1 mentions)
Goat anti mouse igg, by Santa Cruz Biotechnology (1 mentions)
Mouse immunoglobulin g igg, by R&D Systems (1 mentions)
Radio immunoprecipitation assay ripa buffer, by Santa Cruz Biotechnology (1 mentions)
Rhil 8, by R&D Systems (1 mentions)
Dihydrotestosterone, by Merck Group (1 mentions)
Levonorgestrel, by Merck Group (1 mentions)
Nomegestrol acetate, by Merck Group (1 mentions)
Nestorone, by Merck Group (1 mentions)
Gestodene, by Merck Group (1 mentions)
Drospirenone, by Merck Group (1 mentions)
Net a, by Merck Group (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
8 protocols using fulvestrant ici182 780
1
Endocrine Disruptors and Nuclear Receptors
2
Endocrine Disrupting Chemical Screening
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17β-estradiol (E2), 4-nitrophenyl phosphate, diethanolamine, fulvestrant (ICI 182,780), 8-penylnaringenin (8PN), dicofol (Dic), endosulfan (End), fenarimol (Fen), glyphosate (Glp) and methiocarb (Met) were purchased from Sigma-Aldrich (Schnelldorf, Germany), xanthohumol (XN) and iso-xanthohumol (iX) from Extrasynthese (Genay, France). Concerning cell culture, flasks and dishes were obtained from Sarstedt (Nümbrecht, Germany), media (DMEM, DMEM/F-12) as well as fetal bovine serum (FBS) from GIBCO / Thermo Fisher Scientific (Karlsruhe, Germany) and penicillin/streptomycin (P/S) as well as carcoal dextrane-stripped FBS (CD-FBS) from Sigma-Aldrich (Schnelldorf, Germany).
3
Epithelial-Mesenchymal Transition Regulation
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Tissue culture- and fibronectin-coated plates were purchased from Corning (Corning, NY). Recombinant human (rh)FGF-2, rhIL-6, rhIL-8 and rhTGFβ1 and mouse blocking monoclonal antibody to CXCL1/KC (IL-8) and mouse immunoglobulin G (IgG) were purchased from R&D Systems (Minneapolis, MN). H2O2, carbonyl-cyanide m-chlorophenylhydrazzone (CCCP) and Fulvestrant (ICI 182780) were purchased from Sigma-Aldrich (Saint Louis, MO). Radioimmunoprecipitation assay (RIPA) buffer was purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Rabbit monoclonal antibodies to human E-Cadherin (clone 24E10), N-Cadherin (clone D4R1H) XP), Slug (clone C19G7), β-Catenin (clone D10A8) were purchased from Cell Signaling (Danvers, MA). Mouse monoclonal antibody to estrogen receptor α (ERα) (clone F-10), N-cadherin (clone H-4), horseradish peroxidase (HRP)-labeled goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Vybrant CM-DiI fluorescent tracking dye was purchased from Invitrogen-Molecular Probes (Carlsbad, CA).
4
Receptor Binding Assay Protocol
5
Characterization of Immortalized Human Biliary Epithelial Cells
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HiBECs were purchased from tongpai biotechnology co., Ltd, and the cell line was confirmed by the expression of the biliary-type cytokeratins CK19 (using immune fluorescence) (Supplementary Figure 1A) and CK7 (using immunoblotting) (Supplementary Figure 1D) , and the morphology of HiBECs was observed by optical microscope (Supplementary Figures 1B,C) . HiBECs were suspended in RPMI 1640 medium (Invitrogen, California, USA) with 10% fetal bovine serum supplemented with 10 ng/mL of epidermal growth factor (Sigma-Aldrich, Munich, Germany), 5 mg/mL insulin (Sigma-Aldrich, Munich, Germany), 100 units/ml penicillin, and 100 mg/ml streptomycin (Invitrogen, California, USA). When cells reached 60–70% of confluence, 17β-estradiol (Sigma-Aldrich, Munich, Germany), ERα agonist 4,4,4-(4-propyl-[1H] pyrazole-1,3,5-triyl)-tris-phenol (PPT) (Sigma-Alrich, Munich, Germany), ERβ agonist 2,3-bis (4-hydroxyphenyl)-propionitrile (DPN) (Sigma-Aldrich, Munich, Germany) and ERα antagonist fulvestrant (ICI182, 780) (Sigma-Aldrich, Munich, Germany) at various concentrations were added into the medium. The cells were cultured for 48 h and then harvested for further PCR and immunoblotting analyses.
6
Glucocorticoid Receptor Modulation Pathway
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Fluocinolone acetonide (FA), E2, DHT, RU0486 (Mifepristone), and ICI182,780 (Fulvestrant) were from Sigma-Aldrich (St. Louis, MO, USA). Croton oil (CO) was from Santa Cruz Biotechnology (Dallas, TX, USA). We used antibodies against GR (Santa Cruz Biotechnology), ER (EMD Millipore, Danvers MA, USA), REDD1 (Proteintech Group, Rosemont, IL, USA), GAPDH (Sigma-Aldrich), tubulin, and normal IgG (Cell Signaling, San Jose, CA, USA).
7
Ovarian Cancer Cell Line Responses to Estrogen and BPA
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The OVCAR3 and SKOV3 cell lines were a kind gift from Dr Kunle Odunsi (Roswell Park Cancer Institute, Buffalo, New York). The cells were maintained in RPMI/1640 medium supplemented with 100 U/mL of penicillin–streptomycin (Life Technologies, Carlsbad, California) and 10% heat-inactivated fetal bovine serum (FBS; Atlanta Biologicals, Norcross, Georgia). Cells were cultured at 37°C in a 5% CO2 heated incubator.
17β-Estradiol, BPA, and ICI 182,780 (fulvestrant) were purchased from Sigma-Aldrich (St Louis, Missouri). Cells were treated with 1, 5, or 10 nmol/L estradiol or 10, 25, or 50 nmol/L BPA for 24 hours. Where indicated, the ER inhibitor ICI was added at 10 nmol/L. All treatments were for 24 hours, and 95% ethanol (0.1%) was used as the vehicle control.
17β-Estradiol, BPA, and ICI 182,780 (fulvestrant) were purchased from Sigma-Aldrich (St Louis, Missouri). Cells were treated with 1, 5, or 10 nmol/L estradiol or 10, 25, or 50 nmol/L BPA for 24 hours. Where indicated, the ER inhibitor ICI was added at 10 nmol/L. All treatments were for 24 hours, and 95% ethanol (0.1%) was used as the vehicle control.
8
Exploring Receptor Signaling Pathways
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RPMI 1640-GlutaMAX and Phenol-red free RPMI 1640 media were obtained from ThermoFisher Scientific. Charcoal stripped fetal bovine serum was purchased from Atlanta Biologicals. ERα, STAT5a, STAT5b, STAT5, β-actin antibodies and inhibitors of PI3K (Wortmannin) and JAK2 (AG-490) and c-SRC (PP1) were obtained from Santa Cruz Biotechnology (Santa Cruz, CA). Phospho (p)-ERα (Ser118 and Ser167), pSTAT5, pERK1/2, pMEK1/2, ERK1/2, pAKT, pEGFR (Y1068), pEGFR (Y845) and p-Tyrosine antibodies and U1026 (MEK1/2 inhibitor) were purchased from Cell Signaling. ICI 182,780 (Fulvestrant) and PRLR antibody were obtained from Sigma-Aldrich. Human PRL antibody were obtained from National Hormone and Peptide Program, Harbor-UCLA Med. Ctr., Torrance, CA 90502.
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