Incubation:Article Title: Molecular mechanisms underlying AMH elevation in hyperoestrogenic states in males
Article Snippet: .. The DNA-Lipofectamine solutions were added in a 1:2 DNA-Lipofectamine-Optimem (Gibco, 11058021) solution:transfection medium and incubated for 4 h. Transfection medium was then replaced by 10% charcoal-stripped FBS in DMEM, with amino acids, penicillin–streptomycin and amphotericin B, and incubated overnight. .. Cells were incubated with vehicle (ethanol), 17β-oestradiol (E2, Sigma E8875) or the following agonists or antagonists overnight in serum-free medium: (a) ICI 182780, a high affinity ERα and ERβ antagonist (IC50 = 0.29 nM), devoid of any partial agonism both in vitro and in vivo , , (b) PPT (4,4′,4′′-(4-Propyl-[1H]-pyrazol-1,3,5-triyl) trisphenol), a potent and selective ERα agonist with an effect 400-fold greater than on ERβ , (c) MPP (1,3-bis (4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy) phenol]-1H-pyrazoledihydrochloride), a silent, high-affinity selective ERα antagonist, with a 200-fold more powerful effect than on ERβ , (d) G-1 (1-[(3aR*,4S*,9bS*)-4-(6-Bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl]-ethenone), a potent and selective GPER agonist, displaying no activity on ERα and ERβ at 10 μM (Tocris Bioscience) , (e) G-15 [(3aS*,4R*,9bR*)-4-(6-Bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta[c]quinoline], a potent and selective GPER antagonist, displaying no activity on ERα and ERβ at 10 μM (Tocris Bioscience) .
Transfection:Article Title: Molecular mechanisms underlying AMH elevation in hyperoestrogenic states in males
Article Snippet: .. The DNA-Lipofectamine solutions were added in a 1:2 DNA-Lipofectamine-Optimem (Gibco, 11058021) solution:transfection medium and incubated for 4 h. Transfection medium was then replaced by 10% charcoal-stripped FBS in DMEM, with amino acids, penicillin–streptomycin and amphotericin B, and incubated overnight. .. Cells were incubated with vehicle (ethanol), 17β-oestradiol (E2, Sigma E8875) or the following agonists or antagonists overnight in serum-free medium: (a) ICI 182780, a high affinity ERα and ERβ antagonist (IC50 = 0.29 nM), devoid of any partial agonism both in vitro and in vivo , , (b) PPT (4,4′,4′′-(4-Propyl-[1H]-pyrazol-1,3,5-triyl) trisphenol), a potent and selective ERα agonist with an effect 400-fold greater than on ERβ , (c) MPP (1,3-bis (4-hydroxyphenyl)-4-methyl-5-[4-(2-piperidinylethoxy) phenol]-1H-pyrazoledihydrochloride), a silent, high-affinity selective ERα antagonist, with a 200-fold more powerful effect than on ERβ , (d) G-1 (1-[(3aR*,4S*,9bS*)-4-(6-Bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-tetrahydro-3H-cyclopenta[c]quinolin-8-yl]-ethenone), a potent and selective GPER agonist, displaying no activity on ERα and ERβ at 10 μM (Tocris Bioscience) , (e) G-15 [(3aS*,4R*,9bR*)-4-(6-Bromo-1,3-benzodioxol-5-yl)-3a,4,5,9b-3H-cyclopenta[c]quinoline], a potent and selective GPER antagonist, displaying no activity on ERα and ERβ at 10 μM (Tocris Bioscience) .
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