(A) Schematic illustration of the biomarker identification pipeline. We began with the collection of human gallbladder and liver tissues, followed by identification and enrichment analysis of differentially expressed genes. Orthologous genes were then filtered and compared between human and zebrafish. (B) Comparison of upregulated orthologous genes in human and zebrafish (C) comparison of downregulated orthologous genes in human and zebrafish (D) PCA plot of human liver and human gallbladder samples. € (E) PCA plot of zebrafish liver and zebrafish gallbladder samples.

Gene ontology (GO) enrichment analysis of biological processes and KEGG pathway enrichment analysis (A) and (B) are enrichment of upregulated genes. (C) and (D) are enrichment of downregulated genes. (A) and (C) are human GO enrichment. (B) and (D) are Zebrafish GO enrichment. (E) and (F) are enrichment of upregulated genes. (G) and (H) are enrichment of downregulated genes. (E) and (G) are HUMAN KEGG enrichment. (F) and (H) are zebrafish KEGG enrichment.

Relative expression of cep55l, esrp1, nek2, myb, gapdh, and fabp10a in gallbladder and liver. Real-time PCR shows increased mRNA expression of cep55l, esrp1, nek2, and myb genes (n=3) and decreased expression of gapdh and fabp10a (n=3) in gallbladder compared liver tissue, with rpl13a as a housekeeping gene. Values are represented as mean±SEM. Student’s t-test was performed to determine statistical significance. *P<0.05, ****P<0.0001. cep55l, centrosomal protein 55 like; esrp1, epithelial splicing regulatory protein 1, nek2, NIMA related kinase 2; myb, v-myb avian myeloblastosis viral oncogene homolog; gapdh, glycerldehyde-3-phosphatase dehydrogenase; fabp10a, fatty-acid binding protein 10a; rpl13a, Ribosomal protein L13a.