MCF-7 and MDA-MB-231 cells were obtained from the American Type Culture Collection (Manassas, VA). Cells were cultured in DMEM (Invitrogen, Life Technologies, Carlsbad, CA) supplemented with 10% FBS, 2mM L-glutamine, and 1% penicillin-streptomycin-neomycin. MCF-7 cells were stimulated with 1.32 ng/ml phorbol 12-myristate 13-acetate (PMA) (Sigma-Aldrich, St Louis, MO) or 5 ng/ml recombinant TGF-β1 (R&D Systems, Minneapolis, MN) for 60 h, as previously described [28 ]. 0.65 nM triptolide (Santa Cruz, Dallas, TX) and 1 μM NSC 95397 (Santa Cruz) were used for inhibitor experiments. Forward transfection reactions were performed for 6 h with 50 nM human DUSP1 siRNA (sc-35937), DUSP4 siRNA (sc-38998), DUSP6 siRNA (sc-39000), 20 nM PKC-θ siRNA (sc-36252), and 10 nM MOCK siRNA (sc-36869) (Santa Cruz) using Lipofectamine 2000 (Invitrogen).
Triptolide
Manufactured by Santa Cruz Biotechnology
Sourced in United States
Triptolide is a natural compound extracted from the Chinese herb Tripterygium wilfordii. It is a potent immunosuppressant and anti-inflammatory agent with potential applications in research. The core function of Triptolide is to inhibit the transcription of various genes involved in immune and inflammatory responses.
Automatically generated - may contain errors
Lab products found in correlation
Recombinant tgf β1, by R&D Systems (1 mentions)
Nsc 95397, by Santa Cruz Biotechnology (1 mentions)
Pkc θ sirna, by Santa Cruz Biotechnology (1 mentions)
Mock sirna, by Santa Cruz Biotechnology (1 mentions)
Dusp4 sirna, by Santa Cruz Biotechnology (1 mentions)
Mcf 7, by Merck Group (1 mentions)
Lipofectamine 2000, by Thermo Fisher Scientific (1 mentions)
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (1 mentions)
Phorbol 12 myristate 13 acetate pma, by Merck Group (1 mentions)
Mda mb 231, by American Type Culture Collection (1 mentions)
Mcf 7, by American Type Culture Collection (1 mentions)
α minimal essential medium, by Merck Group (1 mentions)
Roswell park memorial institute (rpmi), by Merck Group (1 mentions)
Dulbecco s modified eagle s medium, by Merck Group (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (1 mentions)
Skbr3, by American Type Culture Collection (1 mentions)
Hs578t, by American Type Culture Collection (1 mentions)
Mda mb 231, by National Collection of Authenticated Cell Cultures (1 mentions)
Mcf 7, by National Collection of Authenticated Cell Cultures (1 mentions)
5 protocols using triptolide
1
Modulating Cell Signaling in Breast Cancer
2
Triptolide Effects on MHCC-97H Cells
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Triptolide (molecular formula, C20H24O6; molecular weight, 360.4 g/mol) was purchased from Santa Cruz Biotechnology, Inc. (Hangzhou, China). Triptolide was dissolved in dimethyl sulfoxide (DMSO). MHCC-97H cells were treated with 1 μM, 10 μM, and 25 μM TPL for 72 hours or 25 μM for 2–48 hours.
3
Triptolide Inhibits Breast Cancer Cell Lines
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Triptolide (molecular formula, C20H24O6; molecular weight, 360.4 g/mol) was purchased from Santa Cruz Biotechnology, Inc. (sc-200122; Santa Cruz, CA, USA). Triptolide was dissolved in dimethyl sulfoxide (DMSO). Three breast cancer cell lines, specifically, the highly metastatic MDA-MB-231, HER2-positive BT-474 and ER-positive MCF7 cell lines, were provided by the Department of Oncology at the Hospital of Traditional Chinese Medicine (Yantai, China). MDA-MB-231, BT-474 and MCF-7 cells were cultured in Dulbecco’s modified Eagle’s medium, RPMI and α-minimal essential medium (Sigma-Aldrich, St. Loius, MO, USA), respectively. Cells were cultured at 37°C with 5% CO2 and 100% humidity. The medium was supplemented with 10% fetal bovine serum (FBS; HyClone Laboratories, Inc., Logan, UT, USA), 100 U/ml penicillin and 100 μg/ml streptomycin.
4
Breast Cancer Cell Line Culturing and Triptolide Treatment
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MCF7, MDAMB231, and T47D were obtained from the Type Culture Collection of the Chinese Academy of Sciences (Shanghai, China); SKBR3, Hs578T and BT474 human breast cancer cells were obtained from the American Type Culture Collection (Manassas, VA, USA). Cells were maintained in RPMI-1640 medium (Gibco; Thermo Fisher Scientific, Inc., Waltham, MA, USA) supplemented with 10% fetal bovine serum (Gibco; Thermo Fisher Scientific, Inc.), 10 U/ml penicillin and 10 mg/ml streptomycin. Cells were cultured in a humidified incubator at 37°C with 5% CO2. Triptolide was purchased from Santa Cruz Biotechnology, Inc. (Dallas, TX, USA; cat. no. sc-200122) and dissolved in dimethyl sulfoxide (DMSO). Cells were treated with indicated concentration of Triptolide and cultured in 24, 48 and 72 h and treated with DMSO as control.
5
Dual-Luciferase Assay for PXR Activity
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Dulbecco’s Modified Eagle’s Medium (DMEM) and fetal calf serum (FCS) were purchased from PAN-Biotech (Aidenbach, Germany). Phosphate-buffered saline (PBS) and medium supplements (glutamine, non-essential amino acids, penicillin/streptomycin) were purchased from Sigma-Aldrich (Taufkirchen, Germany). Rifampicin, crystal violet, and dimethylsulfoxide (DMSO) were purchased from Applichem (Darmstadt, Germany) and methanol from Roth (Karlsruhe, Germany). Dovitinib was provided by Sequoia Research Products (Pangbourne, UK). Triptolide was purchased from Santa Cruz Biotechnology (Heidelberg, Germany). Oxaliplatin (dissolved in distilled water) was supplied by the University Hospital’s pharmacy. The Dual-Glo Luciferase Assay System, the pGL4.21 vector, the pGL4.74 [hRluc/TK] Renilla vector, and the FuGene® HD Transfection reagent were purchased from Promega Corporation (Madison, WI, USA). The NR1I2 (NM_003889) human cDNA TrueClone® (pCMV6-XL4 vector, containing the cDNA of the PXR gene NR1I2) was obtained from OriGene (Rockville, MD, USA). Cell culture flasks and white 96-well plates with a white bottom (especially well-suited for luminescence measurements) were obtained from Greiner (Frickenhausen, Germany). The luminescence signal was detected with the SpectraMax iD3 from Molecular Devices (Wokingham, UK).
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