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Fig. 2 | Journal of Experimental & Clinical Cancer Research

Fig. 2

From: JAK/Stat5-mediated subtype-specific lymphocyte antigen 6 complex, locus G6D (LY6G6D) expression drives mismatch repair proficient colorectal cancer

Fig. 2

Intra-tumoral immunophenotypes marked by LY6G6D and FUT4/CD15. a On the top, unsupervised hierarchical cluster of 232 CRC samples (dataset: GSE17536–37) using cell-specific immune-signatures categorized patients into four groups, with distinct cell immune associated gene expression. Data are obtained using the Euclidean distance and Ward linkage method on the matrix of the enrichment scores calculated through ssGSEA. Top tracks represents the expression profile of known immune inhibitory molecules, together with LY6G6D and CD15/FUT4 genes. On the bottom, boxplots of LY6G6D gene expression in each cluster. b Dot plot representing the mean enrichment scores of each immune cell type in any cluster. Color scale represents the positive (red) and negative (blue) enrichment score; dot size indicates the strength of the association. c representative western blot images and quantification of LY6G6D and CD15 expression from CRC samples and matched normal mucosa (n = 12) relative to β-actin used as loading. Data are mean ± standard error of the mean (s.e.m); (n = 3 biological replicates, P* < 0.05, ***P < 0.001, two-tailed Student’s t-test. Low, LY6G6D and CD15 IHC in normal mucosae and tumor specimens; Scale bar, 100 μm. Enlarged is the staining in both malignant cells (T) and stromal (S) immune cells. d Correlation between LY6G6D+ cells, CD8 T-lymphocytes and CD86 staining in CRC samples (five replicates counts, cells mm− 2). Double immunofluorescence from paraffin embedded sections co-stained with antibodies against CD4 (red) and FOXP3 (red) or LY6G6D (green). Scale bar, 50 μm and 20 μm, respectively

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