Fig 1: Transcriptional and TCR profiles of heterogeneous CD8+ TRMs. (A) UMAP plot of heterogeneous CD8+ TRMs clusters. Each point represents a single cell, colored according to the respective cell cluster: C0: red; C1, pink; C2: green; C3: blue; C4: yellow. (B) The bar graph represents the average proportion of different CD8+ TRMs in tumor tissue. C0: red; C1, pink; C2: green; C3: blue; C4: yellow. (C) Dot plot illustrating the Z-score-normalized mean expression (color scale) and percentage of expressing cells (size scale) for specified genes in each CD8+ TRMs cluster. (D) Monocle2 state plot. Each point in the graph represents a cell, and the color represents the state of the cell. The trace shows the cell state change, and the arrow direction indicates the direction of the cell state change. (E) Pseudotime trajectories for different CD8+ TRMs clusters were constructed using Monocle2. Each dot signifies one cell, and each color denotes one cell cluster. The direction of the arrow indicates the progression of the cell state change. (F) Flow cytometry representative plot of differentiation induction experiments detected the conversion of pTRMs to tTRMs after 72 hours of stimulation with IL-2 (100 ng/mL). Above: control group; below: IL-2 treatment group. (G) The bar graphs show the percentages of CD39+ TRMs cells in the control (black) and IL-2 treatment groups (pink) of differentiation induction experiments (n=4). The error bars represent the mean±SD. (H) The heatmap displays area under the curve score t values for the regulation of expression by transcription factors, computed using the SCENIC workflow. Red indicates enriched gene expression; blue indicates decreased gene expression. (I and J) Bar plots showing the numbers of clonotype (I) and clonal cells (J) in each CD8+ TRMs cluster. Different colors represent different clonotypes. Clonotypes were divided into unique (n=1) and clonal (n=2 and n≥3) categories based on cell number. Clonal cells contained at least two cells of the same clonotype. (K) The pie chart showing the proportion of cloned cells and non-cloned cells in each CD8+ TRMs cluster. Different colors represent different groups. Dark yellow represents cloned cells, blue represents non-cloned cells. (L) The pie chart showing the percentage of TCR clones shared between distinct TRMs clusters. Different colors represent different groups. Orange represents proportion of shared clonotypes, blue represents proportion of unshared clonotypes. Numbers indicate the specific proportion of shared clonotypes. (M) The number of TCR clones shared between distinct CD8+TRMs clusters. The intersection relationship between different clusters is represented by points connected by lines; the bar chart in the lower left represents the size of each cluster clone type; the lower right shows the intersection situation of each cluster, and the bar chart above represents the number of TCR intersections in the included clusters. dTRMs, differentiable CD8+ TRMs; IL, interleukin; iTRM, inhibitory CD8+ TRM; nTRM, normal CD8+ TRMs; TCR, T-cell receptor; TRMs, tissue resident memory T cells; tTRM, tumor CD8+ TRMs; UMAP, Uniform Manifold Approximation and Projection.