Fig 2: Protein expression verification in human. (A) Expression levels of lipocalin-2 (LCN2) in colorectal mucosa of GSE11223. (B) Expression levels of LCN2 in blood sample of GSE94648. (C) Immunofluorescence of LCN2 in normal, short-duration ulcerative colitis (UC), long-duration UC, and ulcerative colitis-associated carcinogenesis (UCAC). (D) LCN2 in normal colon tissues and colorectal cancer (CRC) from the Human Protein Atlas (HPA) database. **P < 0.01, ***P < 0.001.

Fig 3: Protein expression verification in animals. (A) Schematic overview of control, acute colitis (AC), chronic colitis (CC) model, and colitis-associated cancer (CAC) model. (B) Weight loss in mouse of control and three modeling groups. (C) H&E staining of colon sections of mouse at week 11. (D) Lipocalin-2 (LCN2) expression level in colorectal mucosa was detected by RT-PCR, Western blot, and ELISA. (E) Immunofluorescence of LCN2 in control, tumor, paracancer, long-duration (LD) ulcerative colitis (UC) model, and short-duration (SD) UC model. (F) Semiquantitative analysis of immunofluorescence images of LCN2. (G) Calprotectin (CALP), myeloperoxidase (MPO), interleukin (IL)-6, and tumor necrosis factor (TNF)-α expression levels in colorectal mucosa and LCN2 in stool were detected by ELISA. **P < 0.01, ***P < 0.001; ns, not significant.

Fig 4: Sox9 regulates the miR-96-5p/Trib3/IL-6 signaling axis to promote kidney injury. a RT-qPCR to detect the expression of Sox9, miR-96-5p, Trib3, and IL-6 in the kidney tissue of each group of mice. b Western blot to detect the expression of Sox9 and Trib3 in the kidney tissue of each group of mice. c kits to detect the levels of serum creatinine (SCr) and urea nitrogen (BUN) in mice. d H&E staining to detect kidney tissue pathology in mice (×200, 50 μm). e Toluidine blue staining to detect inflammation in mouse kidney tissue (×400, 25 μm). f ELISA to detect the expression of TNF-α, IL-1β, and IL-6 in mouse urine. g Immunohistochemistry to detect the expression of KIM-1 and NGAL, indicators related to kidney injury, in mouse kidney tissue (×400, 25 μm); *indicates p < 0.05; *compared with the Sham group, p < 0.05; #compared with the model+sh-NC group, p < 0.05; &compared to a model+sh-Sox9 group, p < 0.05.

Fig 5: Sox9 regulates miR-96-5p to affect LPS-induced kidney injury. a RT-qPCR assay of miR-96-5p expression in kidney tissues of Sham and Model groups of mice. b ChIP assay of Sox9 enrichment in the promoter region of miR-96-5p in HEK-293T cells. c Dual-luciferase reporter gene assay of Sox9 in HEK-293T cells on miR-96-5p transcriptional regulatory activity in HEK-293T cells. d Sox9 and miR-96-5p expression by RT-qPCR after silencing or overexpression of Sox9 in HEK-293T cells. e ELISA to detect the expression of IL-6, TNF-α, and IL-1β in the supernatant of HK-2 cells in Control and LPS groups. f Western blot to detect KIM-1 and NGAL, an index related to kidney injury in HK-2 cells of Control and LPS groups. g RT-qPCR and Western blot to detect the expression of Sox9 and miR-96-5p in HK-2 cells of Control and LPS groups. h RT-qPCR and Western blot for the expression of Sox9 and miR-96-5p in HK-2 cells of each group; *indicates p < 0.05; #indicates p < 0.05.