Aesculetin

Acetonitrile and acetic acid (both of MS grade), methanol (HPLC grade), Folin–Ciocalteu reagent, hydrochloric acid, sodium hypochlorite, and sodium carbonate were purchased from Merck (Darmstadt, Germany). Ultrapure water (Thermofisher Scientific, Bremen, Germany) was used to prepare standard solutions and blanks. Syringe filters (25 mm, nylon membrane, 0.45 μm) were purchased from Supelco (Bellefonte, PA, USA). Analytical standards of phenolic compounds (gallic acid, protocatechuic acid, gentisic acid, p-hydroxybenzoic acid, p-hydroxyphenylacetic acid, caffeic acid, vanillic acid, syringic acid, p-coumaric acid, ferulic acid, aesculin, aesculetin, polydatin, trans-resveratrol, catechin, isoquercetin, myricetin, luteolin, and naringenin) were purchased from Sigma-Aldrich (Steinheim, Germany).

RPMI media 1640 was obtained from the Sigma-Aldrich Chemical (St. Louis, MO, USA), as were all other reagents unless specifically stated otherwise. Fetal bovine serum (FBS), penicillin/streptomycin, and trypsin/EDTA were purchased from the Lonza (Walkersville, MD, USA). IL-8 protein was provided by R&D systems (Minneapolis, MN, USA). Antibodies of α-smooth muscle actin (α-SMA), E-cadherin, fibronectin, vimentin, collagen I, membrane type 1-matrix metalloproteinase (MT1-MMP), tissue inhibitor of metalloproteinase (TIMP)-1, TIMP-2, ZO-1, β-catenin and CXCR2 were purchased from the Santa Cruz Biotechnology (Dallas, TX, USA). Human N-cadherin antibody was supplied by Abcam (Cambridge, UK). Human collagen IV antibody was purchased from Bioss Antibodies (Woburn, MA, USA). Mouse monoclonal β-actin antibody was obtained from Sigma-Aldrich Chemical. Horseradish peroxidase (HRP)-conjugated goat anti-rabbit IgG and goat anti-mouse IgG were purchased from Jackson Immuno-Research Laboratories (West Grove, PA, USA). Essential fatty acid free bovine serum albumin (BSA) and skim milk were supplied by Becton Dickinson Company (Sparks, MD, USA). 4′,6-Diamidino-2-phenylindole (DAPI) was obtained from Santa Cruz Biotechnology.
Aesculetin (Sigma-Aldrich Chemical) was dissolved in dimethyl sulfoxide (DMSO) for live culture with cells; a final culture concentration of DMSO was <0.5 %.

All reagents and solvents used were of analytical grade. Acetonitrile, formic acid (both MS grade), and methanol (HPLC grade) were purchased from Merck (Darmstadt, Germany). Ultra-pure water (Water Purification System, New Human Power I Integrate, Human Corporation, Republic of Korea) was used to prepare standard solutions and blanks. Analytical standards of protocatechuic acid, syringic acid, p-hydroxybenzoic acid, 5-O-caffeoylquinic acid, caffeic acid, aesculetin, isoorientin, p-coumaric acid, quercetin 3-O-glucoside, naringin, luteolin, hispidulin, and isorhamnetin were purchased from Sigma Aldrich (Steinheim, Germany). Standards of deslanoside, lanatoside C, and digoxin were provided by Professor Yang Ye and Dr. Chunping Tang (Shanghai Institute of Materia Medica- SIMM, Chinese Academy of Sciences — CAS, China).

Choline chloride, L-proline, succinic acid, methanol, sodium hydroxide, sodium carbonate, sodium-nitrite, 2,2-Diphenyl-1-picrylhydrazyl (DPPH), trolox, gallic acid, protocatechuic acid, syringic acid, chlorogenic acid, p-hydroxybenzoic acid, aesculetin, caffeic acid, isoorientin, rutin, vitexin, isoquercetin, p-coumaric acid, quercitrin, astragalin, rosmarinic acid, luteolin, quercetin, naringenin, kaempferol, and rutin were purchased from Sigma Aldrich Chemical Co. (St. Louis, MO, USA). Urea, ascorbic acid, and ethyl acetate were purchased from Betahem (Belgrade, Serbia). Lactic acid and tartaric acid were purchased from ICN Biomedicals Inc. (Aurora, OH, USA). Maleic acid and aluminum chloride were purchased from Acros Organics (Geel, Belgium). Folin–Ciocalteu reagent was purchased from Carlo Erba reagents (Milan, Italy). Glycerol was purchased from Zorka Pharma (Šabac, Serbia). All solvents and chemicals were analytical purity grade.