Glutathione monoethyl ester

WT, Orai1^loxp/−, Orai1^CA1ko/−, and Orai2−/− mice on a C57BL/6 background at postnatal days P17 to P22 were used in the experiments. After the mice were deeply anesthetized with CO₂ and decapitated, the brain was immediately removed and immersed in ice-cold slicing solution containing (in mM) 24.7 glucose, 2.48 KCl, 65.47 NaCl, 25.98 NaHCO₃, 105 sucrose, 0.5 CaCl₂, 7 MgCl₂, 1.25 NaH₂PO₄, and 1.7 ascorbic acid (Fluka, Switzerland). The pH value was adjusted with to 7.4 with HCl and stabilized by bubbling with carbogen, which contained 95% O₂ and 5% CO₂, and the osmolality was 290–300 mOsm. Horizontal hippocampal slices 300 μm were cut in the slicing solution by the use of vibratome (VT1200S; Leica, Germany). Brain slices were kept in the recovering solution, which contained (in mM) 2 CaCl₂, 12.5 glucose, 2.5 KCl, 2 MgCl₂, 119 NaCl, 26 NaHCO₃, 1.25 NaH₂PO₄, 2 thiourea (Sigma, Germany), 5 Na-ascorbate (Sigma), 3 Na-pyruvate (Sigma), and 1 glutathione monoethyl ester (Santa Cruz Biotechnology, USA) at room temperature for at least 1 h before the experiment. The pH value of the recovering solution was adjusted to 7.4 with HCl and constantly bubbled with carbogen, and the osmolality was 290 mOsm.

Dulbecco’s Modified Eagle’s Medium (DMEM), trypsin-EDTA, penicillin-streptomycin and dimethyl sulfoxide (DMSO), MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) (M2128), Hoechst 33258 (B1155) and DCF (D6883) were purchased from Sigma–Aldrich (St. Louis, MO, USA). All tissue culture-ware was from Nunc (Roskilde, Denmark). Proteinase inhibitors were from Roche (Madrid, Spain). ECL western blotting substrate was from Pierce (Thermo Fisher Scientific, Rockford, IL, USA). Novex Sharp Pre-Stained Protein Standard (LC5800) (T-3168) were from Invitrogen Life Technologies (Carlsbad, CA, USA). Sorafenib (BAY 43-9006, Nexavar) and Regorafenib (BAY 73-4506, Stivarga) are manufactured by Bayer. Cabozantinib and A-1331852 were purchased from MedChem Express (Monmouth Junction, NJ, USA). Buthionine sulfoximine, MnTBAP chloride and glutathione monoethyl ester were obtained from Santa Cruz Biotechnology (Dallas, TX, USA).

Wild‐type and SEZ6^−/− mice on a C57BL/6 background at postnatal day P15 were used in the experiments. After the mice were deeply anesthetized with CO₂ and decapitated, the brain was immediately removed and immersed in ice‐cold slicing solution containing (in mM) 24.7 glucose, 2.48 KCl, 65.47 NaCl, 25.98 NaHCO₃, 105 sucrose, 0.5 CaCl₂, 7 MgCl₂, 1.25 NaH₂PO₄, and 1.7 ascorbic acid (Fluka, Switzerland).The pH value was adjusted with to 7.4 with HCl and stabilized by bubbling with carbogen which contained 95% O₂ and 5% CO₂, and the osmolality was 290–300 mOsm. 300 μm horizontal hippocampal slices were cut in the slicing solution by the use of a vibratome (VT1200S; Leica, Germany). Brain slices were kept in a recovering solution which contained (in mM) 2 CaCl₂, 12.5 glucose, 2.5 KCl, 2 MgCl₂, 119 NaCl, 26 NaHCO₃, 1.25 NaH₂PO₄, 2 thiourea (Sigma, Germany), 5 Na‐ascorbate (Sigma), 3 Na‐pyruvate (Sigma), and 1 glutathione monoethyl ester (Santa Cruz Biotechnology, USA) at room temperature for at least 1 h before the experiment. The pH value of the recovering solution was adjusted to 7.4 with HCl and constantly bubbled with carbogen, and the osmolality was 290 mOsm.