Potassium hydroxide

glutamine uptake and α-KG production were determined by glutamine and α-KG assay kits (Abcam, Cambridge, UK), respectively. In brief, cultured cells were harvested and then suspended in assay Buffer (Abcam). After that, samples were centrifuged at 9000 rpm for 12 min, and supernatant was collected. Perchloric acid (Abcam) and potassium hydroxide (Abcam) were used to incubate with supernatant, respectively. After performing centrifugation at 12,000 rpm for 12 min, supernatant was collected and analyzed with the microplate reader (Thermo Fisher) with the wavelength at 450 nm for glutamine uptake assay or at 570 nm for α-KG production assay.

The chemicals and reagents used in this study were of an analytical grade standard. Products purchased from British Drug House, England include sulphuric acid, ammonium sulphate, aqua regia (a mixture of HNO₃ and HCl in the ratio of 1:3), anhydrous sodium sulfate, concentrated tetraoxosulphate (VI) acid, nitric acid, silver trioxonitrate, trioxoborate (III) acid, diethyl ether; from May and Baker, England include iron (II) chloride, potassium ferric cyanide, potassium iodide, potassium tetraoxomanganate; from Merck, Germany include ammonium iron (III) sulphate, calcium (III) chloride, sodium hydroxide, ferric chloride, potassium chiocyanate; from Sigma-Aldrich, USA. include acetone, butylalcohol, heptane, hexane, ethanol, methanol, thioglycolic acid, ammonium thiocyanote, bipyridine, diphenol indo 2, 6 –dichlorophenol, potassium iodide, anhydrous sodium sulphate, isopropanol, 4-amino phenol, ammonium hydroxide, glacial acetic acid, hydrogen peroxide, oxalic acid, sodium acetate anhydrous, potassium hydroxide, a-a 1–dipyridyl, hydrocyanic acid, potassium sulfocyanate; from Abcam, USA, include the total aflatoxin ELISA kit (ab285282, type).