Extended Data Fig. 5: Epithelial membrane protein 1 (EMP1) marks HRCs.

a, Scatter plot showing the correlation value between individual genes in the human SMC cohort (x axis) and in mouse primary tumours (y axis) with the EpiHR signature. Genes with correlation scores higher than 0.8 in both datasets are highlighted. b, UMAP of tumour cells from CRC patients in the SMC dataset coloured according to the expression of EpiHR signature (left) and of EMP1 gene (right). c, As in b, for CRC tumour cells from the KUL datasets. d, UMAP representation of Smart-sequencing single cell data of AKTP mouse tumour cells along metastatic relapse sequence coloured by the EpiHR signature (left) and Emp1 gene (right). e, Vector fields representing RNA velocity projected on AKTP primary CRC, micro+small and macro metastases UMAPs, coloured by the pseudotime estimated for each cell with scVelo. f, AKTP tumour cell UMAPs coloured by Emp1 gene expression. g, Smoothed Emp1 gene expression trends fitted with Generalized Additive Models as a function of pseudotime in AKTP primary tumour, micro+small and macro metastases samples. h, UMAP representation of AKP micrometastases coloured by Emp1 gene expression. i, Smoothed Emp1 gene expression trends fitted with Generalized Additive Models as a function of pseudotime in AKP micrometastases samples. j, Representative flow cytometry plot of TOM expression in wild-type and Emp1-iCT AKTP MTOs. k, Relative mRNA expression of indicated marker genes in Emp1-TOM^high and Emp1- TOM^low sorted cell populations from Emp1-iCT AKTP MTOs in vitro. Two-sided t-test after normalizing by Ppia. n = 3 technical replicates. Mean +/- SD. l, Boxplot showing normalized intensity of coreHRC signature expression in Emp1-TOM^high and Emp1- TOM^low cells dissociated from primary tumours 4 weeks post-implantation. Box plots have whiskers of maximum 1.5 times the interquartile range; boxes represent first, second (median) and third quartiles. n = 4 mice per condition. ROAST-GSA adjusted p-values are shown.

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