Morphine hydrochloride was purchased from Sigma and diluted in cell media to 0.25, 0.5, 1, 2, and 4 μM prior to the experiment. Cisplatin, Fluorouracil (5‐FU), paclitaxel, and naloxone were obtained from Selleck. Cisplatin was reconstituted in PBS, and the rest of drugs were reconstituted in dimethyl sulfoxide (DMSO). Cells were exposed to drugs (see figure legends for specific concentrations) for 24 h and then were used in proliferation and apoptosis experiments, and western blot analysis.
Fluorouracil 5 fu
Manufactured by Selleck Chemicals
Sourced in China, United States
Fluorouracil (5-FU) is a synthetic pyrimidine analog used as a pharmaceutical agent. It is a white to off-white crystalline powder. The molecular formula is C4H3FN2O2 and the molecular weight is 130.08 g/mol.
Automatically generated - may contain errors
Lab products found in correlation
Lipofectamine 2000 transfection reagent, by Thermo Fisher Scientific (2 mentions)
Morphine hydrochloride, by Merck Group (1 mentions)
Naloxone, by Selleck Chemicals (1 mentions)
Paclitaxel, by Selleck Chemicals (1 mentions)
Cisplatin, by Selleck Chemicals (1 mentions)
P smad3, by Cell Signaling Technology (1 mentions)
Gapdh, by Cell Signaling Technology (1 mentions)
Bcl 2, by Cell Signaling Technology (1 mentions)
Cleaved caspase 3, by Cell Signaling Technology (1 mentions)
Vimentin, by Cell Signaling Technology (1 mentions)
E cadherin, by Abcam (1 mentions)
Gapdh, by Proteintech (1 mentions)
Ripa lysis buffer, by Applygen (1 mentions)
Tgf β1, by R&D Systems (1 mentions)
Regorafenib bay 73 4506, by Selleck Chemicals (1 mentions)
5 protocols using fluorouracil 5 fu
1
Cytotoxicity Assay with Opioids
2
Investigating Epithelial-Mesenchymal Transition
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PTX (#S1150) and Fluorouracil (5-FU, #S1209) were purchased from Selleck Chemicals LLC (Shanghai, China). Antibodies against OLA1 (also as GTPBP9, #PA5-31227) and Snail (#PA5-11923) were purchased from Invitrogen (San Diego, CA, USA). Antibodies against GAPDH (#5174), Slug (#9585P), Vimentin(#3932S), Smad3 (#9523P), p-Smad3 (#9520P), Smad4 (#9515P), Cleaved caspase3 (#9661), Bax (#2772), and Bcl2 (#2876S) were purchased from Cell Signaling Technology (Danvers, MA, USA). Antibodies against gamma tubulin (#ab11316), Zeb1 (#ab180905) and E-cadherin (#ab76055) were purchased from Abcam (Cambridge, MA, USA). GAPDH was purchased from ProteinTech Inc (Wuhan, China). The second antibodies were purchased from Abbkine Inc (Wuhan, China). SuperEnhanced chemiluminescence (ECL) detection reagents and RIPA lysis buffer were purchased from Applygen Technologies Inc. (Beijing, China). Apoptosis Kit was purchased from KeyGEN BioTECH Inc (Nanjing, China). (4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) were obtained from Solarbio Inc (Beijing, China). All other reagents were from Beyotime (Haimen, China) and Sangon Biotech (Shanghai, China).
3
Colon Cancer Cell Line Culturing
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The DLD1 and HCT116 human colon cancer cell lines and normal fibroblasts (NFs, ATCC® PCS-201-018) were obtained from American Type Culture Collection (ATCC, Manassas, VA, USA) and were cultured according to the vendor’s recommended conditions. MSI-N1014 was synthesized as described previously in a U.S. patent application [12 (link)] (H.S. Huang, D.S. Yu, T.C. Chen, Vol. US Patent No. 8,927,717B1, US, 6 January 2015). Fluorouracil (5-FU) was purchased from SelleckChem (Hsinchu, Taiwan) (cat. no. S1209). Stock solutions of 5-FU and MSI-N1014 were dissolved in 10 mM dimethyl sulfoxide (DMSO; Sigma Aldrich, St. Louis, MO, USA) and kept at −20 °C. The stock solution was further immediately diluted in the sterile medium at the required concentrations.
4
Transfection of miR-15a with 5-FU in TGF-β1 Experiments
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TGFβ1 was purchased from R&D Biosystems, Minneapolis, MN, USA (Cat. 7754-BH) and fluorouracil (5-FU) from Selleck Chemicals, Houston, TX, USA, (Cat. S1209). Control miR and miR-15a were obtained from ThermoFisher Scientific (Cat. AM17111 and AM17100, respectively), while 5-FU-miR-15a was generated by Horizon Discovery (CTM-628208 and CTM-628207). Lipofectamine 2000 Transfection Reagent was purchased from ThermoFisher Scientific (Cat. 11668027) and used according to the manufacturer’s protocol.
5
Regorafenib and Fluorouracil Treatment
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Regorafenib (BAY 73–4506, catalog No. S1178) and Fluorouracil (5-FU, catalog No. S1209) were purchased from SelleckChem. The primary and secondary antibodies used for western blotting and immunohistochemical experiments were all purchased from Cell Signaling Technology unless otherwise specified.
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