Fig. 1: A single-cell and ST profiling of pairwise oral mucosal biopsies, comprising early OSCCs, adjacent precancerous lesions, and matched normal regions.

a Schematic diagram illustrating the workflow of human oral mucosal biopsies processing for scRNA-seq and ST analyses. N normal, DN dysplasia, T tumor. b Representative H&E staining of tissue samples biopsied for scRNA-seq at the distinct stage during OSCC initiation. Scale bar: 100 μm. c H&E images of ST samples. Initiation stages were indicated at different spatial locations under the guidance of a clinical pathologist. d UMAP representation of single cells profiled in the presenting work colored by major cell types (left), tumor initiation stages (right top) and individuals (right, bottom). e Expression levels of diverse marker genes by annotated cell types. f Bar plots showing the proportion of cell types included in each individual for scRNA-seq. g Heatmap displaying enrichment of cell types in ST samples which were annotated in scRNA-seq. Note that the labeled initiation stages of ST samples were carefully divided under the guidance of a clinical pathologist. h ST feature plots of P1 showing expression of marker genes by annotated cell types.