RNAseq

Total RNA was isolated from kidneys using Trizol (Thermo Fisher, 15596018). Samples with RIN > 7.0 (Agilent Bioanalyzer 2100) were used for library construction and sequenced on an Illumina NovaSeq 6000. Reads were mapped to the Mus musculus genome via HISAT2 (v2.2.1) and quantified as fragments per kilobase of transcript per million mapped reads (FPKM). Differentially expressed genes (DEGs) were identified using DESeq2 or edgeR, applying a threshold of fold change ≥2 and false discovery rate (FDR) < 0.05. Gene Set Enrichment Analysis (GSEA v4.1.0) was performed against GO and KEGG databases. Genes were ranked by the Signal2Noise method, with significant enrichment defined by |NES|>1, NOM p-val<0.05, FDR q-val<0.25.