Astrocyte Hypoxia and Sino Treatment

The procedures were performed in accordance with a previously described protocol [28 (link)]. After purification and siRNA transfection as described above, primary astrocytes were cultured with deoxygenated DMEM without glucose and FBS (Gibco, CA, USA) in an incubator (Thermo Scientific) with premixed gas (1 % O₂, 94 % N₂, 5 % CO₂) for 5 h. Then, the cells were given normal DMEM (Gibco, CA, USA) containing 10 % FBS and placed in a CO₂ incubator (95 % air and 5 % CO₂) for 24 h. Cells in the control group were cultured with normal DMEM and 10 % FBS for the same time. After OGD induction, 0.01, 0.1, 1, 10 mM Sino was immediately used to treat astrocytes for 24 h.