Potassium cyanide kcn

Cell culture medium (RPMI-1640) and fetal bovine serum (FBS) were purchased from Biological Industries (Kibbutz Beit Haemek, Israel). Penicillin/streptomycin were obtained from Hyclone, Logan, UT (USA). 7-(1,3-Benzoxazol-2-ylsulfanyl)-3-benzyl-3H-[1,2,3]triazolo [4,5-d]pyrimidine (VAS2870) was purchased from Calbiochem San Diego, CA (USA). Lucigenin was obtained from Roche (Little Falls NJ, USA). Etoposide, 3-(4,5-Dimethyl-2-thiazolyl)-2,5-diphenyl-2H-tetrazolium bromide (MTT), dimethylsulfoxide (DMSO), phorbol myristate acetate (PMA),2,2-diphenyl-1-picrylhydrazyl hydrazide (DPPH), sodium dodecyl sulfate (SDS), quercetin, catechin, 2,3-butanedione monoxime (BDM), taurine, nicotinamide adenine dinucleotide phosphate reduced (NADPH), ethylene glycol bis(β-aminoethylether)-N,N,N’,N’-tetraacetic acid (EGTA), phenylmethylsulfonyl fluoride (PMSF), cytochrome C, potassium cyanide (KCN), and all other chemicals were purchased from Sigma-Aldrich (St. Louis, MO, USA).

Pooled HLMs (Product Number: M0567) from various male human livers were purchased from Sigma-Aldrich company (St. Louis, MO, USA) and stored at −70 °C until use. As stated in the product information sheet: (1) Polymerase chain reaction protocol was used for pathogenicity testing of all liver specimens (male human donners of mixed age and different health states). (2) Each liver was tested to be negative for hepatitis B and C, HIV1 & 2 and HTLV1 & 2 negative. (3) The protein content was 20 mg/mL in 250 mM sucrose to keep HLMs active. Total cytochrome P450 enzymes and FMO activities are mentioned in the product information sheet. Organic solvents are of HPLC grade, and reference powders are of analytical grade. HPLC grade water was prepared inhouse by a Milli-Q plus filtration instrument that was purchased from Millipore company (Burlington, MA, USA). Tepotinib (99.87 %) and NADPH (99.99%) were procured from MedChem. Express Company (Monmouth, NJ, USA). Acetonitrile, formic acid (HCOOH), ammonium formate (NH₄COOH) and potassium cyanide (KCN) were procured from Sigma-Aldrich company (St. Louis, MO, USA).

HPLC grade solvent and analytical grade reference powders were used. Sprague Dawley rats were obtained from Prince Naif bin Abdul Aziz Health Research Center at King Saud University (KSA). Rat liver microsomes (RLMs) were prepared in-house using Sprague Dawley rats.^{28,29 (link)} INF reference powder was purchased from Med Chem Express (Princeton, NJ, USA). Formic acid (HCOOH), acetonitrile (ACN), HPLC-grade glutathione reductase (GSH) and potassium cyanide (KCN) were purchased from Sigma-Aldrich (USA). HPLC grade water was supplied by an in-house Milli-Q plus purification system (USA). Tween-80 was obtained from Eurostar Scientific Ltd (Liverpool, UK). Ethical approval for the animal experiments was obtained from the Animal Ethics Committee of King Saud University (No. KSU-SE-19-52).

Perifusion analyses were performed using Krebs Ringer bicarbonate solution (KRB) containing 0.1% BSA as done previously25 (link), except that 5 mM was substituted for 25 mM sodium bicarbonate. Glucose, streptozotocin, carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone (FCCP), and potassium cyanide (KCN) were all purchased from Sigma-Aldrich (St. Louis, MO). Culture media (RPMI-1640) was supplied by ThermoFisher Scientific (Waltham, MA), and heat-inactivated fetal bovine serum (FBS) was supplied by Atlanta Biologicals (Flowery Branch, GA).

The following reagents (analytical grade) were used in the present investigation without any further purification: Europium(III) nitrate pentahydrate (Eu(NO₃)₃∙5H₂O, Sigma Aldrich, St. Louis, MO, USA, 99.9%), Cerium(III) nitrate hexahydrate (Ce(NO₃)₃∙6H₂O, Sigma Aldrich, St. Louis, MO, USA, 99.9%), Lanthanum nitrate hexahydrate (La(NO₃)₃∙6H₂O, Sigma Aldrich, St. Louis, MO, USA, 99.9%), Titanium (IV) isopropoxide (Ti(OC₃H₇)₄, Sigma Aldrich, St. Louis, MO, USA, 98.0%), Isopropyl alcohol (C₃H₈O, Sigma Aldrich, St. Louis, MO, USA, ≥99.5%), Hydrochloric acid (HCl, Sigma Aldrich, St. Louis, MO, USA, 37.0%), Sodium hydroxide (NaOH, Sigma Aldrich, St. Louis, MO, USA, ≥85.0%), Potassium cyanide (KCN, Sigma Aldrich, St. Louis, MO, USA, ≥97.0%), Picric acid ((O₂N)₃C₆H₂OH, Sigma Aldrich, St. Louis, MO, USA, ≥99.0%), Sodium carbonate (Na₂CO₃, Sigma Aldrich, St. Louis, MO, ≥99.0%), p-Benzoquinone (C₆H₄(=O)₂, Sigma Aldrich, St. Louis, MO, USA, ≥98.0%), Ethanol (CH₃CH₂OH, Sigma Aldrich, St. Louis, MO, USA, 95.0%).

Human thyroid cancer cell lines (FTC133, FTC236, FTC238, BCPAP, and TT) were obtained from Dr. Motoyasu Saji (The Ohio State University) with permission from the researchers who originally established the cell lines. All thyroid cancer cell lines had been tested and authenticated by short tandem repeat profiling analysis to be of thyroid origin. These cell lines express common thyroid oncogenes, including BRAFV600E (BCPAP), loss of PTEN expression (FTC133, FTC236, FTC238), and RETC634W mutation in TT cells.
Cancer cells were propagated in conventional RPMI 1640 medium (Invitrogen, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS), 100 U/mL penicillin, and 100 mg/mL streptomycin in a humidified 5% CO₂ incubator. The cells were sub-cultured with 0.5% trypsin and 0.02% EDTA (Sigma–Aldrich, St. Louis, MO, USA) when the cell confluency reached 80%. All experiments were performed using thyroid cancer cell lines that had been passaged fewer than 20 times. Potassium cyanide (KCN) was purchased from Sigma–Aldrich (Sigma–Aldrich, St. Louis, MO, USA).