Transcriptome sequencing

To investigate the intrinsic mechanisms underlying the effects of L. intestinalis metabolites on the proliferation and migration of IOMM-Lee cells, we performed transcriptomic analysis of IOMM-Lee cells treated with L. intestinalis metabolites (B1 group vs. A group, Fig. 3M). Following RNA sequencing and library construction, protein-coding genes were analyzed for differential expression using DESeq2 software, with thresholds based on fold change and adjusted *p*-values. Volcano, radar plot, and heatmap plots were generated. To investigate the inhibitory effects of ILA on tumor cells in IOMM-Lee + M0 macrophage (PBMC-derived) co-culture systems, we performed eukaryotic reference-based transcriptome sequencing on tumor cells from Group B2 (IOMM-Lee cells in co-culture systems) and Group A (control group), identifying differentially expressed protein-coding genes.