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7 protocols using mpp dihydrochloride

1

Evaluation of Novel Signaling Modulators

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Tamoxifen citrate, ceramide, MPP dihydrochloride, G-1, Gö 6983, Gö 6976, PKCζ-PS and DL-PDMP were all obtained from Tocris Bioscience (Bristol, UK). Myriocin was obtained from Cayman Chemical (Ann Arbor, MI). Unless otherwise noted, all other chemicals were obtained from Sigma Aldrich.
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2

Bisphenol-A and Estrogen Receptor Modulation

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Bisphenol-A was obtained from MP Biomedicals (Cat No 155118; Santa Ana, CA, USA). 17β-oestradiol (Cat No E8875), diazoxide (Cat No D9530), and nifedipine (Cat No D7634) were obtained from Sigma-Aldrich (Saint Louis, MO, USA). SNX-482 (Cat No 4363) was obtained from PeptaNova (Sandhausen, Germany). DPN (Cat No 1494), PPT (Cat No 1426), PHTPP (Cat No 2662), MPP dihydrochloride (Cat No 1991), PD98059 (Cat No 1213), and Wortmannin (Cat No 1232) were obtained from Tocris Cookson (Bristol, UK).
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3

Hippocampal Slice Recordings and Object Location Memory

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For hippocampal slice recording, the compounds were infused into the aCSF bath via an independent perfusion line (6mL/hr). Final aCSF bath concentrations: APV (100 μM; Hello Bio #HB0225), bicuculline (20 μM; Tocris Bioscience #0130), L655,708 (field electrophysiology: 150 nM, whole-cell electrophysiology: 50 nM; Tocris Bioscience #1327), MPP dihydrochloride (3 μM; Tocris Bioscience #1991) were dissolved in DMSO (<0.01%). For the Object Location Memory task, vehicle (0.1% DMSO) or L655,708 (0.5mg/kg in 0.1% DMSO) was injected intraperitoneally 30 min prior to training.
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4

Evaluation of Novel Signaling Modulators

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Tamoxifen citrate, ceramide, MPP dihydrochloride, G-1, Gö 6983, Gö 6976, PKCζ-PS and DL-PDMP were all obtained from Tocris Bioscience (Bristol, UK). Myriocin was obtained from Cayman Chemical (Ann Arbor, MI). Unless otherwise noted, all other chemicals were obtained from Sigma Aldrich.
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5

Compound Acquisition for Research

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Raloxifene hydrochloride, MPP dihydrochloride, and PHTTP were purchased from Tocris Bioscience (Bristol, UK). PMA, micrococcal nuclease from Staphylococcus aureus, the small molecule library used for screening and all other compounds were obtained from Sigma-Aldrich (St. Louis, MO, USA).
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6

Synthesis and Characterization of Bioactive Compounds

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E2 was purchased from Sigma-Aldrich. MPP dihydrochloride (1.3-bis(4-hydroxyphenyl)-4-methyl-5-(4-(2-piperidiny lethoxy)phenol)-1H-pyrazole dihydrochloride); PHTPPP (4-(2-phenyl-5,7-bis(trifluoromethyl)pyrazolo(1,5-a) pyrimidin-3-yl)phenol) and G15 ((3aS*,4R*,9bR*)-4-(6-Bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta[c] quinoline) were purchased from Tocris Biosciences (R&D system).
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7

Selective Estrogen Receptor Antagonists

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The selective ERa antagonist (MPP dihydrochloride), ERb receptor antagonist (PHTPP), and GPR30 receptor antagonist (G15) used in this study were purchased from Tocris Bioscience (Ellisville, MO, USA). E2, DDP, and gemcitabine were purchased from Sigma (St. Louis, MO, USA). The ligands were prepared as 0.1 M stocks in DMSO or 100% ethanol and stored at -20°C. Prior to use, ligands were further diluted into cell culture medium for treatment groups.
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