Fig. 2: NSCLC subclusters and characteristics related to cuproptosis and immune pathways.

a Heatmap of gene expression across 31 pathways: The heatmap displays the expression patterns of three main clusters related to copper-dependent tumor proliferation, immune infiltration, and stromal enrichment across three distinct clusters (Cluster 1, Cluster 2, Cluster 3) in the OAK + POPLAR cohort. The color scale indicates the degree of expression, with blue representing low expression and red representing high expression. Categories such as Treg, B cells, immune response markers, and matrix remodeling are highlighted, providing insight into the differential gene signatures in these groups. b Kaplan–Meier survival analysis: The survival curve compares the progression-free survival (PFS) between the three clusters in the OAK and POPLAR cohort. Cluster 1 exhibits a significantly worse PFS compared to Cluster 2 (p-value < 0.001). c Volcano plot of differential gene expression: This plot shows the differential expression analysis between Cluster 1 and Clusters 2 and 3. Red points indicate significantly upregulated genes, green points indicate significantly downregulated genes, and gray points represent genes with no significant change. d Enrichment analysis: The bar plot shows the top enriched biological processes (BP) identified through Gene Ontology (GO) analysis for upregulated genes in cluster1. The x-axis represents the significance (−log10(FDR)) of the enriched terms, while the y-axis lists the top GO terms. Terms related to skin development, keratinization, and cell communication are highlighted. e Enrichment analysis of downregulated genes in Cluster 1. f Network of differentially expressed genes: a network analysis reveals the complex interactions among differentially expressed genes, highlighting key regulatory pathways. The network nodes represent genes, with edges indicating interactions. Enriched biological processes (BP), molecular functions (MF), and KEGG pathways are shown, underscoring the connectivity between immune response, cell adhesion, and tumor progression pathways. g Gene set enrichment analysis (GSEA): The GSEA plot illustrates the enriched KEGG pathways in the different clusters. Notably, “Cervical Cancer Proliferation” and “Antigen-activated lymphocytes” are among the top enriched terms, with a significant enrichment score. The term’s ranking in the ordered dataset is shown below the plot.