Fig 1: GZMB induced tubular epithelial cells damage in perforin-dependent way. (A) Prf mRNA expression levels in the kidneys of UIRI mouse models (n = 6 biologically independent mice). (B) Detection of Prf in UUO mouse kidneys by qRT-PCR (n = 6 biologically independent mice). (C) Experimental design: HK-2 cells were co-incubated with GZMB (2 μg/mL) and PRF (160 ng/mL), followed by cell viability assessment and transcriptome sequencing analysis. (D) Cell viability monitored by CCK-8 assay. (E) Pearson correlation heatmap between samples. The color scale represents correlation coefficients between samples (closer to 1 indicates stronger correlation). (F) PCA showing transcriptional differences between the GZMB + PRF group (red dots) and NC group (blue dots) (PC1 = 51.21%, PC2 = 21.21%). (G) Volcano plot of DEGs between control and treatment groups. The X-axis represents log2(FC), and the Y-axis represents -log10(FDR). Blue dots: downregulated genes; red dots: upregulated genes; gray dots: non-significant genes. (H) Hierarchical clustering heatmap of DEGs. Each column represents a sample, and each row represents a gene. Colors are scaled based on log10(FPKM+0.000001), with darker shades indicating higher expression. *p < 0.05, **p < 0.01. Data are presented as Mean ± SD. Statistical analysis was performed using two-tailed Student’s unpaired t-test analysis.