The use of animals and protocols were approved by the Laboratory Animal Welfare and Ethics Committee of Southern University of Science and Technology. All reagents used for neuronal cultures were from Thermo Fisher Scientific (USA) unless otherwise specified. Dorsal root ganglia (DRG) neurons were dissected from E13 C57BL/6 mouse embryos in Leibovitz’s L-15 medium and dissociated in TrypLE express enzyme. Dissociated neurons or explants of DRG were plated on acid-washed glass coverslips pre-coated with poly-D-lysine (100 μg/ml, Trevigen) and laminin (3.3 μg/ml, Trevigen), and cultured in Neurobasal medium supplemented with B27 (20 ml/l), GlutaMAX Supplement (2 mM), Penicillin–Streptomycin (100 U/ml, 100 μg/ml), 5-fluoro-2′-deoxyuridine (10 μM, Sigma) and Nerve growth factor (NGF) (50 ng/ml). For bath application of rhein, DRG neurons were plated and cultured for 2 h. Then rhein (Sigma) was added to the medium with final concentration of 50 μM. After 4 h, neurons were either processed for immunostaining or used for axon growth assay by taking phase-contrast images.
Rhein
Manufactured by Merck Group
Sourced in United States
Rhein is a lab equipment product manufactured by Merck Group. It is used for chemical analysis and research purposes. Rhein is a solid compound with specific chemical properties that make it suitable for various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Emodin, by Merck Group (10 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (6 mentions)
Chrysophanol, by Merck Group (5 mentions)
Physcion, by Merck Group (4 mentions)
Aloe emodin, by Merck Group (4 mentions)
Gallic acid, by Merck Group (3 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (3 mentions)
Trizol, by Thermo Fisher Scientific (3 mentions)
Sennoside a, by Merck Group (2 mentions)
Milli q water purification system, by Merck Group (2 mentions)
Dimethyl sulfoxide (dmso), by Merck Group (2 mentions)
Ammonium acetate, by Merck Group (2 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Methanol, by Merck Group (2 mentions)
Cell culture materials, by Thermo Fisher Scientific (1 mentions)
Anti slug antibody, by Cell Signaling Technology (1 mentions)
Phospho gsk3β ser9, by Santa Cruz Biotechnology (1 mentions)
Antibody against hif 1α, by BD (1 mentions)
Goat anti rabbit igg, by Merck Group (1 mentions)
Anti snail, by Cell Signaling Technology (1 mentions)
27 protocols using rhein
1
Rhein Modulates Neuronal Cultures
2
Rhein Antiviability Evaluation on Renal Cells
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Rhein was purchased from Sigma Chemical Company (St Louis, MO, USA). In vitro antiviability effect of Rhein on A498, 786-O and ACHN cells was examined using CCK-8 assay. Cells were plated in 96-well plates (1×104 cells) in 200 μL of full medium each well and then treated with serial dilution of Rhein (0, 10, 20, 40, 80 and 160 μM). After treatment for 48 h, CCK-8 assay was performed using CCK-8 KIT as per the manufacturer’s instructions. Absorbance at 490 nm (OD490) was detected by using a microplate reader.
3
Emodin, AE, and Rhein Modulation of EMT
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Cell culture materials were obtained from Invitrogen (Burlington, Ontario, Canada). The reagents 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide (MTT), propidium iodide (PI) and the PureLink™ HiPure Plasmid DNA Purification Kit were purchased from Sigma (St. Louis, MO, USA). Primary antibodies against PARP and Twist were purchased from GeneTex (Beverly, MA, USA). Anti-HER-2, anti-phospho-Akt (Ser473), anti-E-cadherin, anti-vimentin, anti-Snail, and anti-Slug antibodies were purchased from Cell Signaling (Beverly, MA, USA). Primary antibodies against ILK, phosphor-ILK (Thr173), phospho-mTOR (Ser2448), and phospho-GSK3β (Ser9) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA). Antibodies against YB-1 and β-Actin were purchased from Millipore (Temecula, CA, USA). The antibody against HIF-1α was purchased from BD Biosciences Clontech (San Jose, CA, USA). For Western blotting, the secondary antibodies of horseradish peroxidase (HRP)-conjugated goat anti-mouse IgG and goat anti-rabbit IgG were obtained from Millipore (Temecula, CA, USA), and enhanced chemiluminescence (ECL) reagents were purchased from Sigma-Aldrich. Emodin, AE, and rhein were purchased from Sigma-Aldrich. The RNAi Consortium of YBX-1 was selected by the National RNAi Core Facility.
4
Rhein Effects on Embryo Development
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Zygotes were cultured in 20-μL microdroplets until 99 h post-hCG, at which point pools of 20 compacted embryos were selected, washed, and placed in a new 20-μL microdroplet of KSOMaa supplemented with 0.1% PVA and 0.5% dimethyl sulfoxide (DMSO) (Sigma) (vehicle control) or KSOMaa supplemented with 0.1% PVA, 0.5% DMSO, and 120 μM rhein (Sigma-Aldrich; P/N: R7269). The remaining embryos that failed to develop to the morula stage were designated as degenerate. These were also washed and moved to fresh 20 μL vehicle control microdroplets in cohorts of 20. Unconditioned blank control media microdroplets were set up and similarly cultured for the same time period. At 116 h post-hCG, all embryos were collected, lysed, and stored at −20°C. Fifteen microliters of each of the conditioned culture media and the blank control media were lysed in an equivalent volume of lysis solution and stored at −20°C. This was performed in triplicate.
5
Compounds for Estrogen Receptor Assay
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TSG, emodin, chrysophanol, physcion, rhein, and dimethylsulfoxide (DMSO) and water soluble β-estradiol were purchased from Sigma-Aldrich Co. (St. Louis, MO, USA).
6
Quantification of Anthraquinones in R. palmatum
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Crude R. palmatum extract powder was obtained from Sun Ten Pharmaceutical Co., Ltd. For each tested extract, 1 g of powder was dissolved in 40 ml methanol or distilled water and gently shaken overnight at room temperature. Solution samples were filtered with Whatman No. 1 filter paper, lyophilized in an IWAKI FDR-50P freeze dryer, then sterilized by a 0.44 μm syringe filter and stored at −80 °C until used (100 % stock solution). Also, filtered extract was injected directly into the HPLC instrument with C-18 reverse phase column. Separation was conducted with gradient elution using acetonitrile and 0.1 % phosphoric acid at a flow rate of 1 ml/min, eluent detected at 250 nm. Chrysophanol, rhein, emodin, aloe-emodin, and physcion were purchased from Sigma Chemical Co. of St. Louis, serving as external standards for comparing chromatographic peaks of methanol and water extracts with the retention time of these marker compounds.
7
Rhubarb Compound Efficacy Evaluation
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Ade was obtained from Amresco (Solon, OH, USA) and the fresh Ade solution was prepared daily. The 2% Ade was prepared from dissolving 1 g Ade in 50 mL flour solution. Rhubarb granules were purchased from Tianjiang Pharmacology Co. Ltd (Jiangyin, China) and were dissolved in distilled water to a concentration of 1 g/mL for experimental use. HBSS was purchased from HyClone (Logan, Utah, USA). Rhein, emodin, bafilomycin A1, LiCl, rapamycin, insulin, metformin, Akti, PD098059 and SB203580 were obtained from Sigma-Aldrich Chemical Co. (St Louis, MO, USA).
8
Culturing GC Cells with Rhein and RAD001
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Human GC cells (MGC-803) were kept in our laboratory and were maintained in RPMI 1640 Medium (Gibco, CA, USA) supplemented with 10% fetal bovine serum (FBS; Gibco) at 37°C in 5% CO2 incubator [20 (link)].
Rhein was purchased from Sigma-Aldrich (MO, USA) with a purity ≥ 95%, and RAD001 (Everolimus) was purchased from Sigma-Aldrich. The drugs (Rhein and RAD001) were dissolved in saline.
Rhein was purchased from Sigma-Aldrich (MO, USA) with a purity ≥ 95%, and RAD001 (Everolimus) was purchased from Sigma-Aldrich. The drugs (Rhein and RAD001) were dissolved in saline.
9
Extraction and Authentication of C. fistula Fruits
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Dried and mature C. fistula fruits were collected in May 2016 in the area of Ubon Ratchathani University, Thailand and were authenticated according to the Thai Herbal Pharmacopoeia [30 ] by Asst. Prof. Dr. Bancha Yingngam. The voucher specimen was deposited at the Faculty of Pharmaceutical Science, Ubon Ratchathani University (voucher specimen: BCY UBU No. 028). The woody ripe pod was broken with a knife. The septa with the fruit pulp were separated from the seeds and the shell. The collected plant material was stored in plastic bags in a dark place at 25 °C until use. Rhein from Sigma Chemical Co. (St. Louis, MO, USA) was used as reference substance. All applied organic solvents were of HPLC and analytical grade.
10
Gonadotropin-Induced Cell Death Evaluation
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Rhein, pregnant mare’s serum gonadotropin (PMSG), bovine serum albumin (BSA), 2′,7′-dichlorofluorescin diacetate (DCF-DA) and sodium pyruvate were purchased from Sigma (St. Louis, MO, USA). Human chorionic gonadotropin (hCG) was obtained from Serono (NV Organon Oss, the Netherlands). The TUNEL in situ cell death detection kit was acquired from Roche Molecular Biochemicals (Mannheim, Germany) and CMRL-1066 from Gibco Life Technologies (Grand Island, NY, USA). TRIzol reagent and the reverse transcriptase kit were purchased from Takara Biomedicals (Kusatsu, Japan).
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