RNA-seq of BV2:sample2
BV2 cells were pretreated with siISG15 or negative control (NC) and subjected to OGD for 2 h, followed by 24 h of normal culture. Total RNA was extracted using the NEBNext Ultra RNA Library Prep Kit (Illumina), and libraries were constructed according to the manufacturers instructions. Sequencing was performed on an Illumina HiSeq platform (Wuhan Kangce Technology Co., Ltd..Raw reads were processed with Fastp (v0.23.0) to remove adapters, low-quality reads, and poly-N sequences. Clean paired-end reads were aligned to the mouse reference genome (GRCm39), and transcript abundance was calculated as counts per million. Differentially expressed genes (DEGs) between groups were identified using DESeq2 (v4.4.1), with thresholds of p < 0.05 and |log2 fold change| > 1. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were performed with the ClusterProfiler and org.Mm.eg.db packages, covering biological process, molecular function, and cellular component categories. Gene set enrichment analysis (GSEA) was also conducted using ClusterProfiler, and pathways with adjusted p < 0.05 were considered significantly enriched.