Fig. 1

α-SMA⁺ CAFs correlate with immune infiltration in ESCC. A UMAP visualization of the 13 distinct cell clusters identified from scRNA-seq data (PRJNA777911) from ESCC tumor samples. B Scatter plots showing the correlation between α-SMA⁺ CAFs and immune cell type enrichment score. The relationship was measured by linear regression and non-parametric Spearman correlation. C The t-SNE plot combined from 10 ESCC tumor tissues and overlaid with 25 differently colored phenograph clusters, with each color representing one cluster. D Heat map of the 25 phenograph clusters were determined by normalized median epitome expression of stained markers using IMC antibodies. E The histogram of α-SMA⁺ CAFs, CD163⁺ macrophages (MØs), granzyme B⁺ immune cells, and CD4⁺ and CD8⁺ T cells. Mean intensities for 10 ESCC samples are displayed in different colors. Scatter plots further exhibited the relevance of α-SMA⁺ CAFs to other immune cell types (CD163⁺ MØs, granzyme B⁺ immune cells, CD4⁺ and CD8⁺ T cells) based on the mean intensity. The relation was measured by linear regression and non-parametric Spearman correlation. F/G Representative images of multiplex IF staining of ESCC tumor samples. Scale bar = 100 μm. Scatter plots showing the positive correlation between α-SMA⁺ CAFs and CD163⁺ MØs (n = 12), or between CD163⁺ MØs and immunologic effector cells (CTLA4⁺, FOXP3⁺, CD4⁺ and CD8⁺ cells) (n = 12). P-values were determined by non-parametric Spearman correlation analysis