Transcriptomic analysis of mouse liver in gallstone formation

High-throughput transcriptome sequencing was employed to profile hepatic gene expression in mice. Total RNA was extracted from tissue specimens using TRIzol reagent. Transcriptome libraries were prepared using the Illumina Stranded mRNA Prep kit. mRNA was enriched using oligo (dT) beads, fragmented, and reverse-transcribed. Libraries were sequenced on the NovaSeq platform with paired-end 150 bp reads. Differential gene expression analysis was performed with the DESeq2 package to identify genes with FDR < 0.05 and absolute fold change > 2. GO and KEGG enrichment analyses were conducted to interpret the biological relevance of the DEGs.