Immortalized human embryonic kidney cells (HEK293.T), human microglia (CHME3; a kind gift from prof. dr. M. Tardieu, Universite Paris-Sud, France66 (link)), human oligodendrocytes (MO3.13), mouse neuroblastoma expressing APPswe (N2a/APPswe; a kind gift from prof. dr. T.W. Kim, Colombia University, USA67 (link)), and monkey kidney cells (COS7) were used for in vitro experiments. All cell lines were cultured in DMEM (Sigma-Aldrich) containing 10% heat-inactivated FCS (Invitrogen, Merelbeke, Belgium) and 100 U penicillin/100 µg streptomycin/ml (Invitrogen), at 37 °C/5% CO2. For phytosterol treatment, cells were incubated for 18 hours in culture medium without FCS containing the Eastern plants extracts, brassicasterol (Sigma-Aldrich), β-sitosterol, (Sigma-Aldrich), fucosterol (Sigma-Aldrich), stigmasterol (analytic confirmed purity of 99,9%), phytosterol mix (containing 60% β-sitosterol, 25% campesterol, and 15% stigmasterol; kindly provided by Ingmar Wester Raisio, Finland), T0901317 (Cayman Chemicals, Huissen, the Netherlands), ethanol (VWR), or DMSO (Sigma-Aldrich).
Fucosterol
Manufactured by Merck Group
Sourced in United States
Fucosterol is a type of phytosterol found in various marine organisms, primarily brown algae. It serves as a key functional component in analytical procedures and scientific research applications involving the analysis and characterization of lipids, sterols, and related biomolecules.
Automatically generated - may contain errors
Lab products found in correlation
β sitosterol, by Merck Group (7 mentions)
Campesterol, by Merck Group (4 mentions)
Desmosterol, by Merck Group (4 mentions)
Brassicasterol, by Merck Group (3 mentions)
5α cholestane, by Merck Group (3 mentions)
Stigmasterol, by Merck Group (2 mentions)
Lanosterol, by Merck Group (2 mentions)
Cycloartenol, by Merck Group (2 mentions)
Squalene, by Merck Group (2 mentions)
Lathosterol, by Merck Group (2 mentions)
Zymosterol, by Avanti Polar Lipids (2 mentions)
Penicillin, by Thermo Fisher Scientific (1 mentions)
Streptomycin, by Thermo Fisher Scientific (1 mentions)
Ethanol, by Avantor (1 mentions)
Stigmasterol, by Cayman Chemical (1 mentions)
Fetal calf serum (fcs), by Thermo Fisher Scientific (1 mentions)
β sitosterol, by Cayman Chemical (1 mentions)
T0901317, by Cayman Chemical (1 mentions)
Dulbecco modified eagle medium (dmem), by Merck Group (1 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (1 mentions)
10 protocols using fucosterol
1
Phytosterol Effects on Cell Lines
2
Fluorescence-Based Cellular Assay Protocol
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Fucosterol, propidium iodide (PI), RNase A triton X-100 dimethyl and sulfoxide (DMSO), were obtained from Sigma-Aldrich Co. (St. Louis, MO, USA). All primary and secondary antibodies were purchased from Santa Cruz Biotechnology Inc. (Santa Cruz, CA, USA). The fluorescent probes DCFH-DA, DiOC6, 4′-6-diamidino-2-phenylindole (DAPI), Fetal bovine serum (FBS), RPMI-1640 medium, L-glutamine, antibiotics were obtained from Invitrogen Life Technologies (Carlsbad, CA, USA).
3
Sterol Sulfate Profiling and Synthesis
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Methanol and water, LC/MS grade, were from MercK (Darmstadt, Germany). [25,26,26,26,27,27,27-D7]-cholesterol sulfate standard (CHOS-d) and cholesterol sulfate were purchased from Sigma-Aldrich (Milan, Italy). Reference sterol sulfates were synthesized in-house by chemical derivatization of sterols (24-methylene cholesterol, desmosterol, campesterol, brassicasterol, dihydrobrassicasterol, stigmasterol, fucosterol, and β-sitosterol) that were purchased from Sigma-Aldrich (Milan, Italy). HPLC for synthetic StS standard purification were performed on a JASCO system (PU-2089 Plus quaternary gradient pump equipped with a MD-2018 Plus photodiode array detector and Sedex 85 high-sensitivity LT-ELS detector). UPLC–MS analysis was performed on Q-Exactive hybrid quadrupole-orbitrap mass spectrometer (Thermo Scientific, Waltham, MA, USA) equipped with an Infinity 1290 UHPLC System (Agilent Technologies, Santa Clara, CA, USA).
4
Quantitative GC-MS Analysis of Plant Sterols
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Sterols were silylated with N,O-bis[trimethylsilyltrifluoro-acetamide] (BSTFA), trimethylchlorosilane (TMCS), and pyridine at 70 °C for 1 h. Trimethylsilyl (TMS) derivatives of sterols were analyzed with GC-MS (Shimadzu) equipped with a Phenomenex (USA) ZB-5 Guardian column (30 m × 0.25 mm × 0.25 μm). Sterols were identified using characteristic ions78 (link) and quantified using authentic standard solutions of plant sterol mixture from Larodan (including 53% β-sitosterol, 7% stigmasterol, 26% campesterol, 13% brassicasterol), and cholesterol, desmosterol, ergosterol, and fucosterol from Sigma-Aldrich. The recovery percentage of the sterol samples was calculated using 5-α-cholestane (Sigma-Aldrich) as an internal standard.
5
Multifaceted Antioxidant Analysis Protocol
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5α-cholestane, 37 standard fatty acid methyl ester mixtures, α-tocopherol, γ-tocopherol, δ-tocopherol, fucosterol, standard polyphenol extract, BSTFA + TMCS, campesterol, squalene, 24-Methylenecycloartan-3β-ol, β-sitosterol, 2,2-diphenyl-1-picrylhydrazyl (DPPH), 2,4,6-tris(2-pyridyl)-s-triazine (TPTZ), and 6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid (Trolox) were obtained from Sigma-Aldrich (Saint Louis, MO, USA). All standards had purity ≥ 97%. Folin–Ciocalteu phenol reagent was purchased from Shanghaiyuanye Bio-Technology Co., Ltd. (Shanghai, China). Methanol (LC-MS grade) was purchased from Fisher, USA. Acetonitrile (LC-MS grade) was purchased from Merck KGaA (Darmstadt, Germany). Alkaline microcrystalline cellulose, and other chemicals and reagents of analytical grade, were obtained from Sinopharm Chemical Reagent Co., Ltd. (Beijing, China). Silica dioxide and activated clay were purchased from cNPC Konecranes Technology Co., Ltd. (Wuhan City, Hubei Province, China). All the reagents were directly used without further purification.
6
Formulation of Firefly Luciferase mRNA Lipid Nanoparticles
7
Sterol Profiling by GC-MS
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Cholesterol, stigmasterol, β-sitosterol, 7-dehydrocholesterol, lathosterol (5α-cholest-7-en-3β-ol), squalene, campesterol, brassicasterol, desmosterol, lanosterol, fucosterol, cycloartenol, and 5α-cholestane (internal standard) were acquired from Sigma-Aldrich (Saint-Quentin-Fallavier, France), cycloartanol and cycloeucalenol from Chemfaces (Wuhan, China), and zymosterol from Avanti Polar Lipids (Alabaster, United States). The C7-C40 Saturated Alkanes Standards were acquired from Supelco (Bellefonte, United States). Reagents used for extraction, saponification, and derivation steps were n-hexane, ethyl acetate, acetonitrile, methanol (Carlo ERBA Reagents, Val de Reuil, France), (trimethylsilyl)diazomethane, toluene (Sigma-Aldrich) and N,O-bis(trimethylsilyl)trifluoroacetamide with trimethylcholorosilane [BSTFA:TMCS (99:1); Supelco].
8
Lipid Standards for Biochemical Assays
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Squalene, cholesterol, β-sitosterol, lanosterol, cycloartenol, campesterol, lathosterol, desmosterol, fucosterol, phosphatidylinositol, phosphatidylcholine, phosphatidylethanolamine, phosphatidylserine, phosphatidic acid, lysophosphatidylethanolamine, lysophosphatidylcholine and ceramide phosphorylethanolamine were purchased from Sigma Aldrich (St Louis, MO, USA). Zymosterol was purchased from Avanti Polar Lipids (Alabaster, AL, USA) and ergosterol was purchased from Supelco (Sigma-Aldrich Sweden AB, Stockholm, Sweden). All other growth media components were purchased from Sigma Aldrich (St Louis, MO, USA) and were of microbiological grade.
9
Fucosterol and Anisomycin Modulate Inflammatory and Apoptotic Pathways
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Fucosterol and anisomycin were purchased from Sigma-Aldrich (St. Louis, MO, USA) and dissolved in 2% DMSO. Antibodies purchased from Cell Signaling Technology (Danvers, MA, USA) included TNF-α, PPARγ, RXRα, IL-6, IL-1β, LC3, NF-κB p65, P38 MAPK, p-P38 MAPK, JNK, p-JNK, ERK, and p-ERK. Antibodies purchased from Proteintech (Chicago, IL, USA) included Beclin-1, Bax, Bcl-2, P62, and β-actin. Liver enzymes were detected and analyzed using microplate test kits (Nanjing Jiancheng Biotech, Nanjing, China) and the RNA PCR kit was purchased from Takara Biotechnology (Dalian, China). Enzyme-linked immunosorbent assay (ELISA) kits for TNF-α, IL-6, and IL-1β were acquired from eBioscience (San Diego, CA, USA). A terminal deoxynucleotidyl transferase dUTP nick-end labeling (TUNEL) apoptosis assay kit was purchased from Roche (Basel, Switzerland).
10
Phytosterol Standards Characterization
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The authentic phytosterol standards, including fucosterol, saringosterol, and ostreasterol, were obtained from Sigma-Aldrich (Saint Louis, MO, USA), of which the purity was verified (estimated to be 96.1%, 98.3%, and 98.0% based on 1H NMR data, see Figure S1 ). For 1H NMR measurement, the internal standard (IS) 2,3,4,5-tetrachloronitrobenzene (>98.0%) was purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan), while the solvent deuterated chloroform (CDCl3, >99.8%) was purchased from Kanto Chemical Co., Inc. (Tokyo, Japan). All the other chemicals were of the highest grade available and purchased from Sigma-Aldrich unless otherwise specified.
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