Pups were sacrificed after prior anesthesia 48 h after HI injury (n=6/group) and the cortical tissue in the peri-infarct was obtained. Total RNA was extracted from cerebral cortices and BV2 cells using Trizol Reagent (Invitrogen, Grand Island, NY, USA) and reverse transcribed to cDNA using a cDNA Synthesis Kit (Thermo Fisher Scientific, Waltham, WA, USA). RT-qPCR was performed by the SYBR Green method (K1622, Applied Biosystems, CA, USA). The reaction was performed at 95 ℃ for 5 min followed by 40 cycles of 95 ℃ for 15 s and 55 ℃ for 40 s on an IQ5 PCR machine (Bio-Rad, Hercules, CA, USA). The relative expression of the targeted mRNA was normalized to the expression of β-actin. Sequence-specific primers were designed according to previous literature (20 (link)).