Dimethyl sulfoxide (DMSO), 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), glucose, trichloroacetic acid, thiobarbituric acid, Tris, flavonolignans (silybin, silychristin and silydianin) were all obtained from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Arachidonic acid was purchased from Chrono-Log (Havertown, PA USA). All other chemicals were reagent grade or the highest-quality available.
Silychristin
Manufactured by Merck Group
Sourced in United States
Silychristin is a chemical compound produced by the Merck Group. It is a naturally occurring flavonoid extracted from the milk thistle plant (Silybum marianum). Silychristin serves as a key ingredient in various pharmaceutical and research applications requiring this specific chemical compound.
Automatically generated - may contain errors
Lab products found in correlation
Silybin, by Merck Group (3 mentions)
Silydianin, by Merck Group (2 mentions)
Trichloroacetic acid, by Merck Group (1 mentions)
Thiobarbituric acid, by Merck Group (1 mentions)
Arachidonic acid, by Chrono-Log (1 mentions)
2 amino 2 norbornanecarboxylic acid bch, by Merck Group (1 mentions)
Verapamil, by Merck Group (1 mentions)
Rifampicin, by Merck Group (1 mentions)
Naringin, by Merck Group (1 mentions)
Trypan blue dye, by Bio-Rad (1 mentions)
Jc 1 dye, by Thermo Fisher Scientific (1 mentions)
Cell counter slides, by Bio-Rad (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Rpmi 1640, by Merck Group (1 mentions)
Histopaque 1077, by Merck Group (1 mentions)
Isotype control, by BD (1 mentions)
Cellfix, by BD (1 mentions)
Anti cd61 pe, by BD (1 mentions)
Anti cd61 fitc, by BD (1 mentions)
Anti cd45 pe, by BD (1 mentions)
8 protocols using silychristin
1
Evaluation of Antioxidant Compounds
2
Preparation and Dissolution of Compounds
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2-Amino-2-norbornanecarboxylic acid (BCH), naringin, rifampicin, silychristin, and verapamil were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands) and L-tryptophan from Fluka (Landsmeer, The Netherlands). BCH and verapamil were dissolved in MilliQ water, and naringin and silychristin in dimethyl sulfoxide. L-tryptophan and rifampicin were dissolved in Dulbecco's phosphate-buffered saline (DPBS)/0.1% glucose.
[125I]-3,3′-T2, [125I]-T3 and [125I]-T4 were prepared as previously described (15 (link)).
[125I]-3,3′-T2, [125I]-T3 and [125I]-T4 were prepared as previously described (15 (link)).
3
Flavonolignans Cytotoxicity and Oxidative Stress
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Dimethyl sulfoxide (DMSO), 3-[(3-Cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS), 4-(2-Hydroxyethyl)piperazine-1-ethanesulfonic acid (HEPES), glucose, Tris, NADH, sodium pyruvate, Histopaque®-1077, RPMI 1640 and DMEM mediums as well as the flavonolignans (silybin, silychristin and silydianin), were all obtained from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Trypan blue dye and cell counter slides were sourced from Bio-Rad (Hercules, California, USA). Both the JC-1 Dye and 2′,7′-dichlorodihydrofluorescein diacetate (DCFH-DA Dye), came from Thermo Fisher Scientific (Waltham, MA, USA). All other chemicals were reagent grade or the highest-quality available.
4
Flavonolignans Modulate Platelet Function
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Interleukin-1 beta was purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). Dimethyl sulfoxide (DMSO), Tris and the flavonolignans (silybin, silychristin and silydianin (Figure S1 ) were all obtained from the Sigma-Aldrich Chemical Co. (St. Louis, MO, USA). Flow cytometry reagents: anti-CD61/FITC, anti-CD61/PE, anti-CD45/PE, isotype controls, BD FACSTM Lysing Solution and CellFix were all obtained from Becton Dickinson (San Diego, CA, USA). All of the other chemicals were of reagent grade or the highest quality available.
5
Organotypic Brain Slice Culture with Pharmacological Treatments
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Mouse brains were isolated and transferred into chilled dissection buffer (Hibernate A with 2% B27 supplement [Life Technologies], 2 mM L-glutamine and 1% penicillin/streptomycin) on ice as described (Kim et al., 2013 (link)). Tissue was sectioned as published previously (Kleine Borgmann et al., 2013 (link)) in chilled dissection buffer using a vibratome. Sections (300 μm) were stored in dissection buffer on ice and transferred onto Millicell inserts (Millipore). Organotypic slices were cultured at 37°C and 5% CO2 in a serum-free medium (Kim et al., 2013 (link)) (Neurobasal A [Life Technologies] containing 2% B27 supplement, 2 mM L-glutamine, 1% penicillin/streptomycin, and 80 μM indomethacin [Sigma-Aldrich]). During the entire culture period slices were exposed to 25 μM silychristin (Sigma-Aldrich), 10 μM iopanoic acid (Sigma-Aldrich), 1 mM BCH (R&D Systems), or respective volumes of the solvents DMSO or culture medium as control. Culture medium was replaced every other day and slices were fixed for 1 h in 4% PFA.
6
Iodothyronine Binding Assay Protocol
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Unlabeled iodothyronines, silychristin, bovine serum albumin (BSA), and D-glucose were obtained from Sigma-Aldrich (Zwijndrecht, The Netherlands [NL]). [125I]-T3 and [125I]-T4 were prepared as previously described (23 (link)). X-tremeGENE9 transfection reagent was obtained from Roche Diagnostics (Woerden, NL). Cell culture flasks and plates were obtained from Corning (Schiphol, NL). An overview of the antibodies is provided in Supplemental Table 1 (24 ). Sulfo-NHS-biotin was obtained from Gentaur (Eersel, NL). Neutravidin agarose was obtained from Thermo Fisher Scientific (Bleiswijk, NL).
7
Examining Myofiber Responses to Inhibitors
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Single EDL myofibers were obtained from 2.5- to 4-month-old mice and cultured for 0 hr, 42 hr, or 72 hr (Bentzinger et al., 2013b (link)). In brief, the EDL muscle was carefully removed, only handling the tendons, and digested with 2.5 mL of collagenase I (Sigma; final concentration, 0.2%) in DMEM for 45 min at 37°C under constant agitation. The muscle was then triturated using Pasteur pipettes coated with HS until single fibers came off. Single myofibers were transferred into culture medium (20% FBS in DMEM, 1% chicken embryo extract [Seralab]) in a 24-well plate coated with HS.
For inhibiting MCT8, Silychristin (25 μM, Sigma) was applied to the culture medium of WT and Oatp1c1 KO-derived fiber cultures, whereas control fibers received equal amounts of the solvent (DMSO).
For inhibiting MCT8, Silychristin (25 μM, Sigma) was applied to the culture medium of WT and Oatp1c1 KO-derived fiber cultures, whereas control fibers received equal amounts of the solvent (DMSO).
8
Flavonolignan Extraction and Quantification
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Flavonolignans (silychristin, silydianin, silybin A and B, iso silybin A and B, and taxifolin) were extracted from 1 g ground dry seeds (control, 200 and 600 Gy) or 1 ml cell suspension medium (control, colchicine 0.05%, 200 and 600 Gy) using 80% methanol at 60 ºC for 4 h under vacuum. The extracts were filtered, dried at below 40 ºC and resuspended in 1 mL methanol (Wianowska and Wiśniewski, 2015) . The cell suspension medium was separated from the biomass culture by filtration. For the flavonolignan extraction from the cell suspension medium, samples were taken at day 3 (from the control only) to detect and confirm the production of flavonolignans during the first 3 days, and at days 10 and 12, samples were taken from all the studied treatments plus the control, with three replicates each.
All the standards of flavonolignans (silychristin, silydianin, silybin A and B, iso silybin A and B, and taxifolin) were purchased from Sigma-Aldrich, U.S.A. HPLC-grade methanol and acetic acid were used for the chromatographic assay.
All the standards of flavonolignans (silychristin, silydianin, silybin A and B, iso silybin A and B, and taxifolin) were purchased from Sigma-Aldrich, U.S.A. HPLC-grade methanol and acetic acid were used for the chromatographic assay.
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