Single-cell RNA-seq data of mouse kidneys under Ischemia-reperfusion operation

Male mice aged 8–10 weeks were used to establish the unilateral IR-induced AKI model. Briefly, under pentobarbital sodium (50 mg/kg)-induced anesthesia, mouse flank incisions were made to expose the kidneys, followed by clamping the left kidney pedicle for 30 min to induce ischemic injury. During the surgery, the body temperature of the mouse was maintained at 36.5–37.3°C via a homeothermic pad. The same procedure without renal pedicle clamping was performed in the sham control mice. The mice were sacrificed on day 3 after surgery for further experiments.A single-cell suspension was prepared for library construction and sequencing following an established single-cell RNA sequencing workflow. For cell type identification, gene expression data were processed using the Seurat package. Briefly, the data were normalized using the Normalize Data function and scaled with the Scale Data function. The top 2,000 highly variable genes were selected using the Find Variable Features function and used as input for principal component analysis (PCA). The top 20 principal components were subsequently utilized for cell clustering via the FindClusters function. Dimensionality reduction and visualization were performed using the Uniform Manifold Approximation and Projection (UMAP) method.Differentially expressed genes (DEGs) were identified with the Find Markers function in Seurat, applying the Wilcoxon rank-sum test under default parameters. Genes were considered significantly differentially expressed if they were detected in more than 10% of cells within a cluster and exhibited an average log₂ fold change greater than 0.25.