The RAW 264.7 cells were infected with lentivirus expressing PCP-EZH2, dCas9 and sgRNA-PP7 plasmids. Three days later, pretreated RAW264.7 cells were labeled with 5 μM DiI (Life Technologies). Meanwhile, B16-F10 cells were labeled by incubation with 5 μM DiO (Life Technologies). And then, 10 μM pirarubicin (Sigma) was added in B16-F10 cells for 2 days. The DiI-labeled RAW264.7 cells were scraped from the plate, followed by addition into the plate with DiO-labeled B16-F10 cells. After 4 h co-culture, cells were washed and fixed in 4% PFA (Sigma) for 15 min before labeled nuclei by DAPI (Sigma). Samples were observed by a Nikon A1 Spectral Confocal Microscope (Nikon).
Pirarubicin
Manufactured by Merck Group
Sourced in United States
Pirarubicin is a laboratory reagent used in scientific research. It is a cytotoxic anthracycline compound that functions as a topoisomerase II inhibitor. Pirarubicin is often used in cell culture and biological experiments.
Automatically generated - may contain errors
Lab products found in correlation
A1 spectral confocal microscope, by Nikon (1 mentions)
Paraformaldehyde (pfa), by Merck Group (1 mentions)
4 6 diamidino 2 phenylindole (dapi), by Merck Group (1 mentions)
Horseradish peroxidase hrp conjugated anti mouse igg, by GE Healthcare (1 mentions)
Anti rabbit igg antibody, by GE Healthcare (1 mentions)
Doxorubicin, by Merck Group (1 mentions)
Daunorubicin, by Merck Group (1 mentions)
Idarubicin, by Merck Group (1 mentions)
Recombinant human tnf α, by R&D Systems (1 mentions)
Clean blot ip detection reagent hrp, by Thermo Fisher Scientific (1 mentions)
Verapamil, by Merck Group (1 mentions)
Doxorubicin, by Fresenius (1 mentions)
Curcumin, by Nacalai Tesque (1 mentions)
Resazurin, by Merck Group (1 mentions)
3 4 5 dimethylthiazol 2 yl 2 5 diphenyltetrazolium bromide, by Merck Group (1 mentions)
Rhodamine b, by Merck Group (1 mentions)
Hemin, by Merck Group (1 mentions)
Desferrioxamine mesylate, by Merck Group (1 mentions)
Cyclosporin a, by Novartis (1 mentions)
Vd 550r, by TAITEC (1 mentions)
9 protocols using pirarubicin
1
Lentiviral-mediated Phagocyte-Cancer Cell Interaction
2
Anticancer Agents and Molecular Targets
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Ten milligrams of epirubicin hydrochloride injection “NK” was purchased from Nippon Kayaku and dissolved in normal saline (Otsuka) at the time of use for in vitro and in vivo experiments. Pirarubicin, doxorubicin, daunorubicin and idarubicin were all purchased as hydrochloride salts from Sigma-Aldrich. Recombinant human TNF-α was purchased from R&D Systems. Anti-Foxp3 and anti-GAPDH antibodies were purchased from Abcam. Anti-NFAT1 and anti-NF-κB antibodies were purchased from Cell Signaling Technologies. Anti-Foxp3 antibody for immunoprecipitation was purchased from Santa Cruz Biotechnology. Horseradish peroxidase (HRP)-conjugated anti-mouse IgG and anti-rabbit IgG antibodies were purchased from GE Healthcare. Clean-Blot™ IP Detection Reagent (HRP) was purchased from Thermo Scientific.
3
Curcumin Enhances Doxorubicin Sensitivity in Drug-Resistant Leukemia Cells
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K562 cells and K562/ADR cells were purchased from RIKEN Innovation Co., Ltd. Doxorubicin was from Fresenius Kabi (Bangkok, Thailand) Ltd. Curcumin was purchased from Nacalai Tesque (Kyoto, Japan), pirarubicin (4-Q-tetrahydropyraylDoxorubicin) and verapamil were purchased from Sigma-Aldrich (St. Louis, MO, USA). Rabbit monoclonal anti-P-gp IgG was purchased from Boster Biological Technology, while rabbit polyclonal anti-human GAPDH IgG anti-GAPDH was purchased from Santa Cruz Biotechnology, Santa Cruz, CA, USA. The apoptosis kit was BioLegend (Santa Cruz, CA, USA).
4
Evaluation of Cytotoxic Compounds
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4-hydroxybenzoic acid (4-HBA), 4-hydroxy-3-methoxybenzoic acid (Vanillic acid, VA), Pirarubicin (Pira), Rhodamine B (Rho B), 3-(4, 5-dimethylthiazol-2-yl) 2, 5-diphenyltetrazoliumbromide (MTT), and Resazurin were acquired from Sigma-Aldrich (St Louis, MO, USA).
5
Synthesis of Iron-based Compounds
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Cyclosporin A was purchased from Novartis. Iron(III) chloride anhydrous (FeCl3) was obtained from Fluka; desferrioxamine mesylate, hemin, and Pirarubicin were from Sigma.
6
Pirarubicin-loaded PLGA Nanoparticle Synthesis
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A PLGA NP was generated as described previously18 (link) Briefly, 25 mg of PLGA (#B6017–1 50:50 DL-PLG, Corefront, Inc.) and 4 mg of pirarubicin (Sigma-Aldrich) were dissolved in 1 ml of acetone. The solution was then placed in 20 ml of a 2% polyvinyl alcohol (PVA)/distilled water mixture and irradiated with an ultrasonic wave at a power level of 1.0 (Sonicator 3000, Misonix Inc.) for 10 minutes followed by stirring for 16 h to evaporate the acetone. Then, the supernatant was collected after centrifugation at 200 RCF for 10 minutes and further centrifuged at 15000 RCF for 30 minutes with additional distilled water for washing. The washing step was repeated 3 times, and PBS was added to the pellet and stirred with a homogenizer at 10,000 rpm for 10 minutes to disperse the NPs. Finally, the solution was freeze-dried using a lyophilizer (VD-550R, TAITEC, Tokyo, Japan) for over 48 h. The freeze-dried powder was used for all the experiments. We generated rhodamine-labeled Pir-PLGA NPs the same method by mixing rhodamine with pirarubicin as a landmark of incorporated NPs to AdSCs.
7
Verapamil and Pirarubicin Interaction Assay
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Purified verapamil and pirarubicin were purchased by Sigma-Aldrich (Milan -Italy). Concentrations were determined by diluting stock solutions to approximately 10 -5 M and using ε 480 = 11,500 M -1 cm -1 . Stock solutions were prepared just before use. Buffer solutions were HEPES buffer containing 5 mM HEPES, 132 mM NaCl, 3.5 mM CaCl 2 , 5 mM glucose, at pH 7.3. Rhodamine-123 was purchased by Sigma-Aldrich (Milan -Italy). Cell culture reagents were purchased from Celbio s.r.l. (Milano, Italy). Cul-turePlate 96/wells plates were purchased from PerkinElmer Life Science; Calcein-AM, bisBenzimide H 33342 trihydrochloride were obtained from Sigma-Aldrich (Milan, Italy).
8
Pirarubicin-Induced Oxidative Damage Assay
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Materials. Pirarubicin, superoxide dismutase (SOD; 3,000 U/mg from bovine erythrocytes), catalase (45,000 U/mg from bovine liver) and cytochrome c (from equine heart) were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Plasmid DNA (pBR322) and DNA gel loading dye (6×) were from Toyobo Co. (Osaka, Japan). Copper chloride (CuCl 2 •2H 2 O) was from Nacalai Tesque Co (Kyoto, Japan). Diethylenetriamine-N,N,N',N'',N''-penta-acetic acid (DTPA) and bathocuproinedisulfonic acid were from Dojin Chemicals Co. (Kumamoto, Japan). 3-(Methylthio) propionaldehyde (methional) was from Tokyo Kasei Co. (Tokyo, Japan). All other chemicals used were of the highest purity commercially available.
9
Oxidative Stress Resistance in Leukemia Cells
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Materials. Pirarubicin was purchased from Sigma Chemical Co. (St. Louis, MO, USA). Proteinase K was obtained from Merck (Darmstadt, Germany). Fluorescent probes, 2',7'-Dichlorofluorescin diacetate (DCFH-DA) and 3,3'-dihexyloxacarbocyanine iodide [DiOC 6 (3) ] were purchased from Molecular Probes (Eugene, OR, USA). All other chemicals used were of the highest purity commercially available.
Cell culture and treatment with THP. HP100 cells have been derived from human leukemia HL-60 cells by repeated exposure to H 2 O 2 , followed by outgrowth of viable cells. HP100 cells were approximately 340-fold more resistant to H 2 O 2 than HL-60 cells (15) . Catalase activity of HP100 cells is 18-times higher than that of HL-60 cells (16) . When scavengers such as catalase, superoxide dismutase and reduced glutathione are added in cell culture medium, these scavengers exist in the extracellular environment (17, 18) .
Cell culture and treatment with THP. HP100 cells have been derived from human leukemia HL-60 cells by repeated exposure to H 2 O 2 , followed by outgrowth of viable cells. HP100 cells were approximately 340-fold more resistant to H 2 O 2 than HL-60 cells (15) . Catalase activity of HP100 cells is 18-times higher than that of HL-60 cells (16) . When scavengers such as catalase, superoxide dismutase and reduced glutathione are added in cell culture medium, these scavengers exist in the extracellular environment (17, 18) .
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