Fig 1: Identification and validation of macrophage polarization in chronic allograft fibrosis. (A) volcano plot of differentiated expressed genes (DGEs) in GSE25902; (B-C) Gene set enrichment analysis of DEGs in GSE25902 in GO (B) and KEGG (C) databases; (D) immune cell infiltration results of DEGs using Cibersort software. The red bars represent the relative abundance of specific immune cells in the CAD group, and the blue bars represent the relative abundance of corresponding immune cells in the stable group; (E) HE and Masson staining results in renal specimens from healthy volunteers (n = 5) and patients with chronic allograft dysfunction (CAD; n = 5); (F-G) Indirect double immunofluorescence staining of CD68(+) iNOS(+) M1 macrophages (F) and CD68(+) CD206(+) M2 macrophages (G) in Normal (n = 5) and CAD (n = 5) groups; (H-L) PCR analysis of CD68 mRNA (H), iNOS mRNA (I), CD86 mRNA (J), Arg-1 mRNA (K), and Fizz1 mRNA (L) in Normal (n = 5) and CAD (n = 5) groups. **p < 0.01 compared to Normal group; ***p < 0.001 compared to Normal group; ****p < 0.0001 compared to Normal group. Each experiment was duplicated at least thrice.