Astrocytic prostaglandin transporter SLCO2A1 protects blood-brain barrier integrity following ischemic stroke

Total RNA from naïve astrocytes, astrocytes treated by OGD, astrocytes treated by ODG and Shco2a1-shRNA was used as input for RNA sequencing library preparation utilizing the KCTM Digital mRNA Library Prep Kit (Seqhealth Tech. Co., Ltd., Wuhan, China), according to the manufacturer's instructions. The enriched libraries were quantified and sequenced on a DNBSEQ-T7 platform (MGI) using the PE150 sequencing mode to generate paired-end reads. Briefly, mRNA was enriched and cleaved into short fragments using a fragmentation buffer, followed by reverse transcription into cDNA using random primers. The cDNA fragments were purified and end repaired, and a poly(A) tail was added, followed by ligation with Illumina sequencing adapters. The ligation products were then size selected by agarose gel electrophoresis, amplified by PCR, and sequenced using the DNBSEQ T7. DEGs were assessed by analysis of differential RNA expression between 3 groups. Transcripts with the parameter of FDR below 0.05 and an absolute fold change of 2 or greater were considered differentially expressed.