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3 protocols using exemestane

1

Activating murine PAR2 receptor

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Exemestane, letrozole and anastrozole were purchased from Tocris Bioscience (Bristol, UK). The activating peptide (PAR2-AP, SLIGRL-NH2) and its reverse peptide (PAR2-RP, LRGILS-NH2) of the murine PAR2 receptor were synthesized from G. Cirino (University of Naples, Naples, Italy) and dissolved in distilled water. If not otherwise indicated, all other reagents were from Sigma-Aldrich (Milan, Italy). HC-030031 was synthesized as previously described45 (link).
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2

Compound Acquisition and Experimental Setup

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Compounds were obtained as follows: sulforaphane (Sigma-Aldrich), exemestane (Tocris), letrozole (Cayman), (+)-JQ-1 and (−)-JQ-1 (MedChemExpress), tamoxifen (Sigma-Aldrich), 4-OHT (Sigma-Aldrich), hematoxylin (Sigma-Aldrich), eosin Y (Sigma-Aldrich), osmium tetroxide (Sigma-Aldrich), Fluo-4 AM (Invitrogen), Fura-2 AM (Invitrogen), ascorbate (Sigma-Aldrich), AM-0902 (Tocris), olive oil (Sigma-Aldrich), corn oil (Sigma-Aldrich), Dulbecco’s modified Eagle’s medium (DMEM; Gibco), fetal bovine serum (Sigma-Aldrich), penicillin-streptomycin (Gibco), l-glutamine (Gibco), forskolin (Sigma-Aldrich), collagenase/dispase (Sigma-Aldrich), Hanks’ balanced salt solution (HBSS; Gibco), laminin (Roche), poly-d-lysine (Sigma-Aldrich), protease inhibitor cocktail (Sigma-Aldrich), and 10% normal goat serum (Thermo Fisher Scientific).
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3

Comprehensive Chemical Reagent Inventory

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Chemicals were purchased as follows: Sigma-Aldrich (St. Louis, MO): 17β-estradiol (E2), cycloheximide (CHX, translation inhibitor), 4-hydroxytamoxifen (4-OHT), finasteride (5α-reductase inhibitor), miconazole (general P450 inhibitor), exemestane (aromatase inhibitor), STX64 (steroid sulfatase inhibitor), 5-fluorouracil (5-FU), doxorubicin (Dox), actinomycin D (ActD, a transcriptional inhibitor), and flutamide (a selective androgen receptor modulator (SARM)); Tocris (Ellisville, MO): Fulvestrant (ICI 182, 780), 2,3-bis(4-hydroxyphenyl)propionitrile (DPN), an ERβ-selective agonist; 4,4',4”-(4-Propyl-[1H]-pyrazole-1,3,5-triyl)trisphenol (PPT), an ERα-selective agonist; and 4-[2-Phenyl-5,7-bis(trifluoromethyl)pyrazolo[1,5-a]pyrimidin-3-yl]phenol (PHTPP, an ERβ-selective inhibitor); Steraloids (Wilton, NH): dehydroepiandrosterone (DHEA), dihydrotestosterone (DHT), DHEA 3β-sulfate (DHEA-S), 5-androstene-3α,17α-diol (ADIOL), 5-androstene-3,17-dione (ADIONE), and 5α-androstane-3β,17β-diol (3β-Adiol). The SARM bicalutamide (Casodex) was generously provided by Astra Zeneca (Macclesfield, UK).
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