“…After administering pentobarbital sodium (50 mg/kg, intraperitoneally, CAS, 57-33-0, Xiya Reagent, Chengdu, China), the brain was quickly removed, and the cortex and hippocampus were separated on ice, following which 0.01M PBS (10 μL/mg) was added to the samples, which were then cut using scissors on ice. Sequentially, the cut issue was filtered through a stainless steel mesh (200 mesh/inch) for the single-cell suspensions ( Cheng et al, 2021 ). In accordance with the instructions of the ROS Assay Kit (Beyotime, S0033, Shanghai, China), 500 μL DCFH-DA diluted with serum-free culture medium (1:1000, 10 μM/L) were added to the single cell suspensions (250 μL) for 40 min incubation at 37°C without daylight.…”