Nanodrop 2100 spectrophotometer

Cortical penumbra tissue and cell samples were lysed with TRIzol reagent (Invitrogen), total RNA was extracted according to the manufacturer’s protocol, and the quality of the RNA was assessed with a Nanodrop 2100 spectrophotometer (Agilent). Reverse transcription from RNA to cDNA was performed with the PrimeScript RT regent kit (Vazyme, Nanjing, China, R323-01). Quantitative real-time PCR was performed in a Roche LightCycler^® 96 system (Roche, Mannheim, Germany) with a SYBR green kit (Accurate, Wuhan, China, AG11701). The primers are as follows:
CXCL3: forward primer- CCAGACAGAAGTCATAGCCAC, reverse primer-CTTCATCATGGTGAGGGGCTT;
CXCL11: forward primer- GGCTTCCTTATGTTCAAACAGGG; reverse primer- GCCGTTACTCGGGTAAATTACA;
CCL20: forward primer- GCCTCTCGTACATACAGACGC, reverse primer- CCAGTTCTGCTTTGGATCAGC;
CCL22: forward primer- AGGTCCCTATGGTGCCAATGT; reverse primer- CGGCAGGATTTTGAGGTCCA;
IL12a: forward primer- CTGTGCCTTGGTAGCATCTATG; reverse primer- GCAGAGTCTCGCCATTATGATTC;
IL23a: forward primer- ATGCTGGATTGCAGAGCAGTA; reverse primer- ACGGGGCACATTATTTTTAGTCT;
Serpine1: forward primer- TTCAGCCCTTGCTTGCCTC; reverse primer- ACACTTTTACTCCGAAGTCGGT;
GAPDH: forward primer- AGGTCGGTGTGAACGGATTTG, reverse primer-TGTAGACCATGTAGTTGAGGTCA;
β-actin: forward primer- GGCTGTATTCCCCTCCATCG, reverse primer- CCAGTTGGTAACAATGCCATGT.

Cortical arterioles and cell samples were lysed with TRIzol reagent (Invitrogen). Total RNA was isolated according to the standard protocol provided by Invitrogen and the quality of the RNA was assessed with a Nanodrop 2100 spectrophotometer (Agilent). Then, the RNA was reverse-transcribed into cDNA with a PrimeScript RT reagent Kit (Takara, #RR037A). Quantitative real-time PCR was performed on an ABI 7500 PCR instrument (Applied Biosystems) with a SYBR green Kit (Takara, #DRR820A). The mRNA expression in each sample was calculated by the 2^-(ΔΔCt) method after normalization to the expression of the internal control GAPDH. The primer sequences are shown in the Table S1.