Gefitinib, Dioscin, Triptolide, and Nature product library were obtained from Selleckchem (Houston, TX). All other chemicals were acquired from Sigma Chemical (St. Louis, MO). Anti-EGFR, anti-total AKT, anti- phosphorylated (p) -AKT, anti-total ERK, and anti-(p)-ERK antibodies were purchased from Cell Signaling (Danvers, MA). Anti-SHP2 antibodies were obtained from Genetex (Irvine CA). All other antibodies were purchased from Santa Cruz Biotechnology (Dallas, TX).
Triptolide
Manufactured by Selleck Chemicals
Sourced in United States
Triptolide is a natural compound extracted from the plant Tripterygium wilfordii. It is a chemical reagent commonly used in laboratory research settings. Triptolide exhibits diverse biological activities and has been studied for its potential therapeutic applications.
Automatically generated - may contain errors
Lab products found in correlation
Melphalan, by Merck Group (2 mentions)
Spironolactone, by Selleck Chemicals (2 mentions)
Bendamustine, by Merck Group (2 mentions)
4 hydroperoxy cyclophosphamide, by Santa Cruz Biotechnology (2 mentions)
Pyr 41, by Merck Group (2 mentions)
Gefitinib, by Selleck Chemicals (1 mentions)
Anti phosphorylated akt, by Cell Signaling Technology (1 mentions)
Dioscin, by Selleck Chemicals (1 mentions)
Anti total erk, by Cell Signaling Technology (1 mentions)
Anti p erk, by Cell Signaling Technology (1 mentions)
Anti total akt, by Cell Signaling Technology (1 mentions)
Anti egfr, by Cell Signaling Technology (1 mentions)
Pfi 3, by Selleck Chemicals (1 mentions)
α amanitin, by Merck Group (1 mentions)
Mec 1 cells, by Leibniz Institute DSMZ (1 mentions)
Flavopiridol, by Selleck Chemicals (1 mentions)
Plad b, by Bio-Techne (1 mentions)
Phorbol myristate acetate (pma), by Merck Group (1 mentions)
Decitabine, by Selleck Chemicals (1 mentions)
Sunitinib, by Selleck Chemicals (1 mentions)
11 protocols using triptolide
1
Molecular Pathway Inhibitor Screening
2
Small Molecule Modulation of Embryonic Development
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α-Amanitin was purchased from Sigma (A2263-1 mg) and dissolved in 1 ml ddH2O. For injection, a 1:20 solution was freshly prepared in ddH2O, and 2 nl (0.2 ng) was injected into embryos at one-cell stage. Triptolide (PG490- 1 mg, Selleckchem) was diluted in dimethyl sulfoxide (DMSO) to obtain a 2.5 mM stock. For drug treatment, the stock was further diluted to 10 mM in embryo medium. Manually dechorionated embryos were reared in the Triptolide solution from the 128-cell stage until 4.3 hpf. PFI-3 (catalog no. S7315-5 mg, Selleckchem) was dissolved in DMSO to a 5 mM stock solution. For drug treatment, dilutions of PFI-3 stock in ddH2O (1–100 mM) were freshly prepared, thoroughly vortexed, and 1–2 nl was injected into one-cell stage embryos.
3
Investigating HSF1 in Chronic Lymphocytic Leukemia
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Triptolide and C646 were purchased from Selleck chemicals. HSF1 expression plasmid (HSF1-GFPN3) was obtained from Addgene. Mec-1 cells were purchased from DSMZ. Mec-2 and WAC3-CD5+ cells were obtained from Dr. H. Shi (GRU Cancer Center, GA). All cells were cultured as described previously and authenticated by surface staining and flow cytometry for CD19, CD20 and CD79a [53 (link)]. Stable knockdown of HSF1 was carried out in Mec-1 and WaC3-CD5+ cells using lentiviral non-targeted or two independent HSF1 shRNA constructs (Sigma Aldrich, MO). De-identified and de-linked primary CLL samples were obtained from the Biorepository Core Facility of Kansas University Medical Center, after informed consent using an institutional review board-approved protocol (HSC-5929). CD19+ B cells from newly diagnosed, relapsed or treatment refractory CLL patients were isolated from the samples utilizing a magnetic CD19-positive selection kit (Stem Cell Technologies, Vancouver, BC), as described previously [54 (link)]. The purity of the isolated CD19+ B-cell fraction was assessed using CD19-PE conjugated antibodies (BD Biosciences, San Jose, CA) and flow cytometry. Positively selected cells were re-suspended in 20% FBS containing RPMI prior to performing the studies described.
4
Cell Treatment with Chemical Modulators
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PMA (Sigma-Aldrich, P8139) was added to media at 25 ng ml−1 at 37 °C with 5% CO2 for denoted time. JQ1 (Tocris, 4499) was added to media at 0.5 μM at 37 °C with 5% CO2 for 4 h unless denoted otherwise. Triptolide (Tocris, 3253; Selleckchem, S3604) was added to media at 1 μM at 37 °C with 5% CO2 for 4 h unless denoted otherwise. Flavopiridol (Selleckchem, S1230) was added to media at 1 μM at 37 °C with 5% CO2 for 1 h unless denoted otherwise. Dox (MP Biomedicals, 198955) was added to media at 1 μg ml−1 at 37 °C with 5% CO2 for 24 h unless denoted otherwise. The splicing inhibitor, Plad B (ref. 54 (link); Tocris, 6070), was added to media at 5 μM at 37 °C with 5% CO2 for 2 h unless noted otherwise.
5
Cytotoxic Agents Procurement Protocol
6
Cytotoxic Compounds Acquisition Protocol
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Melphalan, bendamustine and PYR41 were purchased from Sigma (St Louis, MO, USA), spironolactone and triptolide from Selleck Chemicals LLC (Houston, TX, USA) and 4-hydroperoxycyclophosphamide from Santa Cruz Biotechnology (Dallas, TX, USA). Melflufen was obtained from Oncopeptides AB (Stockholm, Sweden).
7
Renal Cell Carcinoma Cell Line Culturing
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The human RCC cell lines (OS-RC-2, Caki-2, Caki-1, A498, 786-O, ACHN, 769-P, and KETR-3) and HK-2 cells were obtained from the Chinese Academy of Sciences (Shanghai, China). A498 and ACHN cells were incubated in Minimum Essential Medium (MEM) (10-010-CV, Corning, USA) supplemented with 10% foetal bovine serum (FBS, 16000044, Gibco, USA), and the other cells were incubated in Roswell Park Memorial Institute (RPMI) 1640 (10-040-CV, Corning, USA) containing 10% FBS. Cells were grown as a monolayer on plastic cell culture dishes at 37°C in a humidified atmosphere containing 5% CO2. Sunitinib, decitabine, triptolide and SCH772984 were purchased from Selleck Chemicals (China). MG132 and cycloheximide (CHX) were purchased from APExBIO (USA). Lonafarnib and betulinic acid (BetA) were purchased from TargetMol USA.
8
Cancer Cell Lines Transfection Assay
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Human liver cancer cell lines (Huh7, SMMC‐7721, QGY‐7703, HepG2) and LO2 normal human hepatic cells were obtained from Guangzhou Jennio Biotech Co., Ltd (Guangdong, China) and cultured in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum. The cells were transfected with a control vector or DDX1 overexpression plasmids. In some experiments, recombinant human IFN‐γ (100 ng/mL; InvivoGen, CA, USA) and NF‐κB inhibitor (triptolide, 15 μmol/L; Selleck Chemicals, Houston, USA) were added to the culture systems.
9
Notch Inhibitor Screening in Cells and Zebrafish
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Natural compounds (#L1400) and FDA-approved chemicals (#L1300), including triptolide and DAPT for the Notch inhibitor screening test, were purchased from Selleckchem, (Houston, TX, USA). DMSO was purchased from Corning (USA, Manassas, VA, USA). Pyridone 6 (P6) was purchased from R&D Systems (a Bio-Techne Brand, Minneapolis, MN, USA). In cell-based experiments, P6, DAPT, P6 + DAPT, TP, or DMSO were treated for a selective concentration of 50 nM, 100 nM, 200 nM, 500 nM. The concentrations of each signal inhibitor and TP were determined through referencing their use in various cells [39 (link),40 (link),41 (link)] and zebrafish [74 (link),75 (link)]. The concentration ranges typically used in cell-based experiments to test anticancer agents was applied [76 (link)]. They were incubated at 37 °C, 5% CO2, for 24 h. To reduce error, chemicals were diluted with Opti-MEM (Welgene, Republic of Korea) before suspension. In zebrafish experiments, 24 hpf embryos were placed in 90ϕ Petri dishes filled with 10 mL of E3 buffer (with 0.001 M methylene blue). Embryos were treated with DMSO for the negative control or a 500 nM concentration of P6, DAPT, P6 + DAPT, or triptolide. They were incubated for 72 h in an incubator at 28.5 °C.
10
Evaluating Pharmacological Inhibitors
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NVP-2, Nutlin-3a, 5-fluorouridine, 5-fluorodeoxyuridine, THZ531 and Pictilisib were from MedChemExpress. 5-fluorouracil, Senexin A, OTS964 and Triptolide were from Selleck Chemicals. YLK-5–124 and THAL-SNS-032 were a gift from Nathanael S. Gray’s Laboratory (Stanford University, USA). iCDK9 was a gift from Qiang Zhou’s Laboratory (University of California, Berkley, USA). PP2A activator DT-061 was a gift from Jukka Westermarck’s Laboratory (University of Turku, Finland).
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