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6 protocols using matairesinol

1

Osteoclastogenesis Assay with M-CSF and RANKL

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Penicillin, streptomycin, cell culture medium, and fetal bovine serum (FBS) were purchased from Invitrogen Life Technologies. Mouse soluble macrophage-colony stimulating factor (M-CSF) and RANKL were purchased from R&D Systems. The CCK-8 assay kit was purchased from Dojindo Molecular Technologies. Antibodies against nuclear factor of activated T cells (NFAT)c1, c-Fos, and actin were purchased from Santa Cruz Biotechnology and Antibodies against MAP kinases from Cell Signaling Technology. Matairesinol was purchased from Sigma-Aldrich and dissolved in DMSO (dimethylsulfoxide; Sigma-Aldrich).
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2

Quantification of Lignans in Materials

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Gallic acid, hydroxymatairesinol, isolariciresinol, lariciresinol, matairesinol, pinoresinol, secoisolariciresinol, and secoisolariciresinol glucoside were provided by Sigma-Aldrich (Milan, Italy). Formic acid (HCOOH) and HPLC-MS solvents such as methanol (MeOH) and water (H2O) were from Sigma-Aldrich (Milan, Italy).
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3

Antioxidant Compound Extraction Protocol

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ABTS (2,2′-azinobis-(3-ethylbenzthiazoline-6-sulfonic acid)) (CAS: 30931-67-0) and DPPH (2,2-diphenyl-1-picrylhydrazyl) (CAS: 1898-66-4) were purchased from Sigma-Aldrich® (Darmstadt, Germany); gallic acid, potassium peroxodisulfate and Na2CO3 were purchased from Fluka (Darmstadt, Germany); methanol was purchased from Carlo Erba (Milan, Italy); and Folin–Ciocalteu (FC) reagent was purchased from Merck (Darmstadt, Germany). The solvents used for the high-performance liquid chromatography (HPLC) were of HPLC-grade purity; methanol, water and acetonitrile were obtained from J. T. Baker (Gliwice, Poland) and trifluoroacetic acid was obtained from Fisher Scientific (Loughborough, U.K.). Reference standard compounds were used for confirmation of identity and HPLC quantification: matairesinol (CAS: 580-72-3), (+)-isolariciresinol (CAS: 549-29-8) and (−)-hydroxymatairesinol (CAS: 347359-71-1) were obtained from Sigma-Aldrich® (Darmstadt, Germany); secoisolariciresinol (CAS: 145265-02-7) was obtained from Fluka (Darmstadt, Germany); and (+)-lariciresinol (CAS: 27003-73-2) and (+)-pinoresinol (CAS: 487-36-5) were obtained from PhytoLab (Dutendorfer, Germany).
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4

Standards for Phenolic Compound Analysis

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Commercial
standards of quercetin, quercetin-3-glucuronide, trans-resveratrol, trans-resveratrol-glucoside, (−)-epicatechin
and dimers B1 and B2, and 3-O-glucosides of cyanidin,
delphinidin, malvidin, peonidin, petunidin, isorhamnetin, and syringetin
were purchased from Extrasynthese (Genay, France). (+)-Catechin, p-hydroxybenzoic acid, 3,4-dihydroxybenzoic acid (protocatechuic
acid), p-coumaric acid, gallic acid, caffeic acid,
ferulic acid, vanillic acid, syringic acid, matairesinol, and secoisolariciresinol
were acquired from Sigma-Aldrich (St. Louis). Caftaric acid and kaempferol-3-glucoside
were purchased from Purifa-Cymit (Barcelona, Spain). Naringerin and
coutaric acid were purchased from Fluochem (Hadfield, England) and
Phytolab (Madrid, Spain), respectively. The solvents, methanol (HPLC
grade), acetonitrile (HPLC-MS grade), and formic acid (HPLC grade),
were purchased from Scharlab Chemie (Sentmenat, Catalonia, Spain).
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5

Matairesinol and 5-FU Interaction Study

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Matairesinol and 5-fluorouracil (5-FU) were both purchased from Sigma-Aldrich (St. Louis, MO, USA). Both chemicals were dissolved in dimethyl sulfoxide (DMSO). Information regarding the antibodies used in our study is presented in Table 1.
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6

Phytochemicals in Cell Cycle Regulation

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Cyanidin-3-glucoside, cyanidin-3-rutinoside, acetic acid, butyric acid, ellagic acid, propinoic acid, protocatechuic acid, valeric acid, matairesinol, lariciresinol, pinoresinol, secoisolariciresinol, and other chemicals were purchased from Sigma-Aldrich (St. Louis, MO). Fructooligosaccharides, xylooligosaccharides, and galactooligosaccharides were obtained from Creative Dynamics (Shirley, NY). Charcoal/dextran-treated fetal bovine serum (FBS) was procured from HyClone (Logan, Utah). Antibodies against cyclin-B1, cyclin-E2, p21, phospho-cdc2, Myt1, CDC25B, and β-actin were obtained from Cell Signaling Technology (Danvers, MA). Primers for RT-qPCR and siRNA for estrogen receptor-α (ERα) were purchased from Thermo Fisher Scientific (Grand Island, NY).
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