Fig. 2: Characterization of cellular heterogeneity, cuproptosis-related gene expression, and pathway enrichment in low- and high-grade LUAD.

A t-SNE plots of single-cell transcriptomes derived from tumors classified as high-grade (left) or low-grade (right). Each point represents an individual cell, colored by its assigned cell type. B Violin plots comparing cuproptosis-related gene expression levels across the indicated cell populations in low-grade versus high-grade tumors. Significance levels are denoted (***p < 0.001). C, D Gene Set Enrichment Analysis (GSEA) of high-grade vs. low-grade groups, showing enrichment profiles for metabolic (Glycolysis and Proteasome) and stress-associated (Cellular response to stress, Cellular response to starvation, Cellular response to hypoxia and Cellular response to external stimuli) pathways. Black vertical lines in the enrichment plots indicate leading edge genes contributing to the enrichment signals. E Second-level t-SNE clustering of epithelial cells, revealing distinct subpopulations within the epithelial compartment. F t-SNE visualization comparing the distribution of epithelial subclusters in high-grade (left) and low-grade (right) tumors. Stacked bar plots showing the proportion of major cell types across individual tumor samples (G) and high- vs. low-grade groups (H).