Erastin2 and ML162 were synthesized at Acme Bioscience. Nutlin-3 (Cat. no. S1061), nutlin-3b (Cat. no. S8065), gemcitabine HCL (Cat. no. S1149), and MI-773 (Cat. no. S7649) were purchased from Selleck Chemicals. Pyrazofurin (Cat. no. SML1502), HU (Cat. no. H8627), MPA (Cat. no. M5255), ferrostatin-1 (Cat. no. SML0583), Doxycycline hyclate (Cat. no. D9891), and L-BSO (Cat. no. B2515) were purchased from Sigma-Aldrich. BSO and gemcitabine HCL were dissolved directly into cell media. C11-BODIPY 581/591 was prepared as a stock solution in methanol and stored before use at −20°C. All other compounds were prepared as stock solutions in DMSO and stored before use at −20°C.
L bso
Manufactured by Merck Group
Sourced in Germany, United States
The L-BSO is a laboratory equipment product from Merck Group. It is designed for the measurement of biological samples. The core function of the L-BSO is to provide accurate and reliable data for scientific research and analysis.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by Corning (2 mentions)
Ferrostatin 1, by Merck Group (1 mentions)
Doxycycline hyclate, by Merck Group (1 mentions)
Mi773, by Selleck Chemicals (1 mentions)
Pyrazofurin, by Merck Group (1 mentions)
L 733 060, by Bio-Techne (1 mentions)
K sfm, by Thermo Fisher Scientific (1 mentions)
L buthionine sulfoximine, by Merck Group (1 mentions)
Sulfasalazine, by Merck Group (1 mentions)
Tung oil, by Merck Group (1 mentions)
N hydroxysuccinimide nhs 130672, by Merck Group (1 mentions)
Hgcl2, by Fujifilm (1 mentions)
C57bl 6j, by CLEA Japan (1 mentions)
C57bl 6j mice, by CLEA Japan (1 mentions)
Paraffin, by Sakura Finetek (1 mentions)
Hematoxylin, by Sakura Finetek (1 mentions)
6 protocols using l bso
1
Small Molecule Compound Preparation and Storage
2
Hyperosmotic Stress-Induced Apoptosis in Corneal Epithelial Cells
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Mouse corneal epithelial cell line (TKE2) was presented from Dr. Tetsuya Kawakita of Keio University (Tokyo, Japan) and cultured in keratinocyte serum-free medium (KSFM, Invitrogen, Carlsbad, CA) [39 (link)]. To induce apoptosis caused by hyperosmotic stress, TKE2 cells were incubated overnight in bovine pituitary extract (BPE)-free KSFM and subsequently exposed to hypertonic media (450, 550 and 650 mOsm) achieved by addition of glucose for 12–48 h. To detect the inhibiting effects on hyperosmotic stress-induced apoptosis, 0.1, 1 or 10 μM SP (Calbiochem, San Diego, CA) was pre-incubated 2 h before the treatment of high glucose. In some experiments, TKE2 cells were treated with the NK-1 receptor antagonist (1 μM L-733,060, Tocris, Minneapolis, MN), the AKT inhibitor V (40 μM Triciribine, Calbiochem, San Diego, CA), or glutathione depleting agent L-Buthionine-sulfoximine (100 μM, L-BSO, Sigma-Aldrich, St. Louis, MO) in the presence or absence of substance P.
3
Orthotopic Xenograft Tumor Model Treatment
4
Orthotopic Xenograft Murine Model for Breast Cancer
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This study was reviewed and approved by the Fox Chase Cancer Center Institutional Animal Care and Use Committee (Protocol #16-10) and complies with ethical regulations for animal research. Mice were housed in a dedicated laboratory animal facility with 12-h light:dark cycle, at 70 F +/−2 degrees, and 40–70% relative humidity. Orthotopic xenografts were generated by implanting 2.5 million MDA-MB-231 cells in 100 µL phosphate-buffered saline (PBS) mixed with 100 µL growth factor-reduced Matrigel (Corning) bilaterally into the fourth inguinal fat pad of four- to six-week-old female NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. After 14 days, when tumors were roughly 50 mm3, animals were randomized into treatment groups. Mice were treated on weekdays with either safflower oil (Whole Foods 365) and Tung oil (Sigma-Aldrich 440337) at a dose of 100 µL administered by oral gavage. L-BSO (Sigma) was administered via the drinking water (20 mM) ad libitum, as previously reported23 (link),83 (link).
5
Folic Acid-Targeted Dendrimer-Encapsulated Fluorescein
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Polyurea (PURE) dendrimers were synthesized following our supercritical-assisted polymerization methodology [22 (link)]. FA-NHS and PUREG4-FA2 were synthesized following our reported protocol [31 (link)]. All chemicals and solvents were used as received without further purification. Folic acid (FA) and N,N-dicyclohexylcarbodiimide (DCC; A10973) (99% purity) were obtained from Alfa Aesar (Kandel, Germany). Fluorescein (FL; F7505), Triethylamine (TEA; 471283) (≥99.5% purity), N-hydroxysuccinimide (NHS; 130672) (98% Purity) and L-buthionine sulfoximine (l -BSO: B2515) (≥97% purity) were obtained from Sigma-Aldrich (Darmstadt, Germany).
The encapsulation of FL in PUREG4-FA2 (FL@PUREG4-FA2) followed the same methodology used for L-BSO encapsulation [31 (link)]. Typically, in a vial, FL (0.0131 mmol, 5.3 mg) was dissolved in 1 mL of distilled water. To this solution, the folate-target dendrimer (PUREG4-FA2) (6.46 µmol, 56.6 mg) was added. The mixture was then left overnight at RT, in the dark, and under stirring. After this period, the product was purified by dialysis (MWCO 100–500 Da) and characterized by 1H NMR.
The encapsulation of FL in PUREG4-FA2 (FL@PUREG4-FA2) followed the same methodology used for L-BSO encapsulation [31 (link)]. Typically, in a vial, FL (0.0131 mmol, 5.3 mg) was dissolved in 1 mL of distilled water. To this solution, the folate-target dendrimer (PUREG4-FA2) (6.46 µmol, 56.6 mg) was added. The mixture was then left overnight at RT, in the dark, and under stirring. After this period, the product was purified by dialysis (MWCO 100–500 Da) and characterized by 1H NMR.
6
Metallothionein Knockout Mouse Model
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MT-I/II null mice with null mutations of the MT-I and MT-II genes, and wild-type mice were kindly provided by Dr. K.H.A. Choo (Murdoch Institute for Research into Birth Defects, Royal Children's Hospital, Parkville, Australia) (Michalska and Choo, 1993) and were of a mixed genetic background of 129 Ola and C57BL/6 strains. F1 hybrid mice were mated with C57BL/6J mice (CLEA Japan, Tokyo, Japan) and their offspring were backcrossed to C57BL/6J for six generations. Both MT-I/ II null mice and wild-type mice were generated by mating with heterozygous (MT+/-) mice. The mice were routinely bred in the vivarium of the National Institute for Environmental Studies, reproducing normally and displaying no overt abnormalities related to physical state or behavior. Both strains of mice were housed in cages in ventilated animal rooms with a controlled temperature of 23 ± 1°C, a relative humidity of 55 ± 10%, and a 12 hr light/dark cycle. They were maintained on standard laboratory chow and tap water ad libitum, and received humane care throughout the experiment according to the guidelines of the National Institute for Environmental Studies.
L-BSO was purchased from Sigma-Aldrich (St. Louis, MO, USA). Paraffin, hematoxylin, and eosin were procured from Sakura Finetek Japan (Tokyo, Japan). HgCl 2 and other chemicals were purchased from Wako Pure Chemical Industries (Osaka, Japan).
L-BSO was purchased from Sigma-Aldrich (St. Louis, MO, USA). Paraffin, hematoxylin, and eosin were procured from Sakura Finetek Japan (Tokyo, Japan). HgCl 2 and other chemicals were purchased from Wako Pure Chemical Industries (Osaka, Japan).
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