Fig. 5: Validation of intratumoral heterogeneity and CD44-related 1C metabolism pathways in human intestinal-type gastric cancer via single-cell transcriptomics.

A Schematic illustration of single-cell RNA sequencing public data consisting of two subtypes of human gastric cancer tissue, diffuse- and intestinal-type. B, C UMAP plots displayed nine individual clusters from 195,263 single cells with three types of samples, diffuse, intestinal, and normal. Annotation was done through ‘SingleR’ package. D Dotplot showing CD44 expression levels across diffuse- and intestinal-type gastric cancers, as well as normal samples. E UMAP of diffuse- and intestinal-type clusters subsetted from (C), with a total of 107,389 cells. F UMAP of epithelial cell type clusters, derived from (E), encompassing a total of 12,351 cells with 12 clusters. G UMAP of epithelial cell clusters from the intestinal-type clusters, extracted from (F), comprising a total of 3898 cells with seven clusters. H Vlnplot showing CD44 expression levels across seven clusters of intestinal epithelial cell types. I UMAP of clusters categorized into ‘CD44_high’ and ‘CD44_low’ expressing groups as shown in (H). J Vlnplot depicting CD44 expression levels of clusters categorized into ‘CD44_high’ and ‘CD44_low’ groups. K Vlnplot illustrating the expression levels of 1C metabolism-related genes in ‘CD44_high’ and ‘CD44_low’ expressing groups. L Vlnplot demonstrating the enrichment levels of 1C metabolism-related KEGG pathways in the ‘CD44_high’ and ‘CD44_low’ expressing groups. GSEA KEGG pathway analysis was performed using the ‘escape’ package. Statistical comparisons were performed using two-tailed Student’s t-test (*P < 0.05, **P < 0.01, ***P < 0.001, ****P < 0.0001, ns not significant).