Fig 2: UCHL3 was highly expressed in liver cancer. A: The expression level of UCHL3 in different types of tumors. The heatmap depicted the ratio of UCHL3 expression between cancer and normal tissues. ACC Adrenocortical carcinoma, BLCA Bladder Urothelial Carcinoma, BRCA Breast invasive carcinoma, CESC Cervical squamous cell carcinoma and endocervical adenocarcinoma, CHOL Cholangio carcinoma, COAD Colon adenocarcinoma, DLBC Lymphoid Neoplasm Diffuse Large B-cell Lymphoma, ESCA Esophageal carcinoma, GBM Glioblastoma multiforme, HNSC Head and Neck squamous cell carcinoma, KICH Kidney Chromophobe, KIRC Kidney renal clear cell carcinoma, KIRP Kidney renal papillary cell carcinoma, LAML Acute Myeloid Leukemia, LGG Brain Lower Grade Glioma, LIHC Liver hepatocellular carcinoma, LUAD Lung adenocarcinoma, LUSC Lung squamous cell carcinoma, MESO Mesothelioma, OV Ovarian serous cystadenocarcinoma, PAAD Pancreatic adenocarcinoma, PCPG Pheochromocytoma and Paraganglioma, PRAD Prostate adenocarcinoma, READ Rectum adenocarcinoma, SARC Sarcoma, SKCM Skin Cutaneous Melanoma, STAD Stomach adenocarcinoma, TGCT Testicular Germ Cell Tumors, THCA Thyroid carcinoma, THYM Thymoma, UCEC Uterine Corpus Endometrial Carcinoma, UCS Uterine Carcinosarcoma, UVM Uveal Melanoma. B: The expression level of UCHL3 in normal liver tissues and liver tumor tissues. C: The level of UCHL3 mRNA in liver tumor at different grades. D: The expression of UCHL3 in normal liver tissues, primary cancer and cancer that have spread to lymph nodes. E: Immunohistochemistry (IHC) staining of UCHL3 in human liver tumors at different grades. T tumor, N normal tissue. Scale bar: 50μm. F: The methylation level of UCHL3 promoter in normal liver tissues and liver tumor tissues. G: The overall survival rate of patients with UCHL3 high-expression and low-expression liver cancer. Bar, SD

Fig 3: UCHL3 facilitated the progression of cell cycle. A GO analysis of differentially expressed genes in shUCHL3 cell. B The expression of Cyclin B1, Cyclin A2, Cyclin D1, MAD2, Caspase 3 and UCHL3 in control and UCHL3 knockdown HepG2 cells. C Cell apoptosis in control and UCHL3 knockdown cells was analyzed by flow cytometry. D Ki67 staining in control and UCHL3 knocked-down HepG2 cells. Scale bar: 10μm. E: The quantification of D. Data were presented as the mean ± SD of three independent experiments. *P < 0.05

Fig 4: UCHL3 regulated cell metabolism. The effects of UCHL3 in ribose phosphate metabolic process (A), starch and sucrose metabolism (B), glycolysis/gluconeogenesis (C), biosynthesis of amino acids (D), phenylalanine metabolism (E) and steroid hormone biosynthesis (F)

Fig 5: Knocking down UCHL3 triggered autophagy in liver cancer cell. A The expression of LC3, P62 and TFEB in control and shUCHL3 cells. B Immunostaining of LC3 in control and shUCHL3 cells. Scale bar: 10μm. C The survival rate of control and shUCHL3 cells treated with different concentration of Chloroquine (CQ). Data were presented as the mean ± SD of three independent experiments. D The effects of CQ on the expression of Cyclin B1, Cyclin A2, and MAD2 in control and shUCHL3 cells. E Ki67 staining in control and shUCHL3 cells treated with DMSO and CQ respectively. Scale bar: 10μm