Quercetin que

Quercetin (QUE; Sigma-Aldrich, Inc., St. Louis, MO, USA), oxaliplatin (OXP; LC Laboratories, Woburn, MA, USA), and sulforaphane (SFN; LKT Laboratories, Inc., St. Paul, MN, USA) were dissolved in dimethyl sulfoxide (DMSO; Sigma-Aldrich, Inc.) before use. Unless otherwise stated, all chemicals were obtained from Sigma-Aldrich, Inc.

C2C12 myoblasts were seeded on culture plates in PM to achieve a 90% confluence, then the medium was changed to the basal differentiation medium (DM), supplemented with 20 µM epicatechin (EC, Sigma-Aldrich, Burlington, MA, USA), 10 µM genistein (GEN, Sigma-Aldrich), 10 µM luteolin (LUT, Sigma-Aldrich), 50 µM quercetin (QUE, Sigma-Aldrich), or 10 µM naringenin (NAR, Sigma-Aldrich) to induce differentiation. DM was DMEM containing 2% horse serum (HS; Thermo Scientific, Waltham, MA, USA) and 1% P/S. The differentiation lasted for 3 days, and the medium was changed every day.
PSCs were seeded on Matrigel-coated plates and cultured in PM for 2–3 days. When cells reached 90% confluence, differentiation was induced in DM supplemented with NAR at the indicated concentration for 5–7 days. In some experiments, ERβ antagonist PHTPP (MedChemExpress, Shanghai, China) at 10 μM was added to DM alone or in combination with NAR. The medium was changed every day.

Quercetin (Que) (purity ≥ 95%, HPLC), cobalt(II) perchlorate hexahydrate (purity ≥ 98%), 4-morpholinepropanesulfonic acid (MOPS) (purity ≥ 99.5%, titration), dimethyl sulfoxide for spectroscopy (DMSO) (purity ≥ 99.8%), and poly(acrylic acid sodium salt) (PAA) (average Mw ~5100, GPC, purity ≥ 99%) were purchased from Sigma-Aldrich and used without purification. Stock solutions of Que were prepared in pure DMSO and then properly diluted in a buffered aqueous solution (pH 7.4, MOPS 10 mM) to have a final organic solvent content of only 5% v/v. Buffer solutions were prepared by weighing proper amounts of MOPS and titrating with a sodium hydroxide solution to the desired pH. All Que solutions were always freshly prepared and stored in the dark to avoid possible light-induced alterations. The cobalt(II) stock solution was prepared by dissolving the corresponding perchlorate salt in water and titrating the resulting solution with standard EDTA, using orange xylenol as an indicator [60 ]. Poly(acrylic acid) stock solutions were prepared at pH 7.4 in MOPS 10 mM; the concentration of the polymer was expressed per monomer unit of acrylic acid. High-purity water (Millipore, Milli-Q Element A 10 ultrapure water) and A-grade glassware were employed throughout.