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A) UMAP projection of SMART-Seq data showing clustering of tumor-resident hybrid cells (TrHCs, EpCAM⁺/ECAD⁺/CD45⁺), circulating hybrid cells (CHCs, EpCAM⁺/ECAD⁺/CD45⁺), tumor cells (EpCAM⁺/ECAD⁺), and immune cells (CD45⁺) from patient-matched colorectal tumor and peripheral blood samples (n = 3 patients; n=3 PBMC samples, n=2 tumor samples). B) UMAP colored by CHC subclusters (CHC1–3) with distinct transcriptional states. C) UMAP colored by patient ID. D) Dot plot showing the top five differentially expressed genes defining each major cell type. E) Gene set enrichment analysis of gene ontology - biological processes highlighting functional differences among CHC1–3 clusters. F) UMAP of 10X Genomics data enriched for tumor-resident hybrids and circulating hybrid cells, colored by hybrid signature scores derived from SMART-seq CHC cluster modules. G) UMAP of the 10X Genomics dataset annotated by major cell types, showing distribution of hybrid-enriched populations. H) Aneuploidy prediction in 10X Genomics dataset distinguishing diploid versus aneuploid populations. I) Violin plots of RUNX1 genes identified in tumor-macrophage murine hybrids ( RUNX1, THBS1, SPI1, GRN, PF4, LGALS3 ). ( p = *** <0.001, **<0.01, *<0.05)

Journal: bioRxiv

Article Title: Cell fusion reprograms tumor cells and promotes RUNX1-mediated invasion and dissemination in colorectal cancer

doi: 10.64898/2026.04.12.717781

Figure Lengend Snippet: A) UMAP projection of SMART-Seq data showing clustering of tumor-resident hybrid cells (TrHCs, EpCAM⁺/ECAD⁺/CD45⁺), circulating hybrid cells (CHCs, EpCAM⁺/ECAD⁺/CD45⁺), tumor cells (EpCAM⁺/ECAD⁺), and immune cells (CD45⁺) from patient-matched colorectal tumor and peripheral blood samples (n = 3 patients; n=3 PBMC samples, n=2 tumor samples). B) UMAP colored by CHC subclusters (CHC1–3) with distinct transcriptional states. C) UMAP colored by patient ID. D) Dot plot showing the top five differentially expressed genes defining each major cell type. E) Gene set enrichment analysis of gene ontology - biological processes highlighting functional differences among CHC1–3 clusters. F) UMAP of 10X Genomics data enriched for tumor-resident hybrids and circulating hybrid cells, colored by hybrid signature scores derived from SMART-seq CHC cluster modules. G) UMAP of the 10X Genomics dataset annotated by major cell types, showing distribution of hybrid-enriched populations. H) Aneuploidy prediction in 10X Genomics dataset distinguishing diploid versus aneuploid populations. I) Violin plots of RUNX1 genes identified in tumor-macrophage murine hybrids ( RUNX1, THBS1, SPI1, GRN, PF4, LGALS3 ). ( p = *** <0.001, **<0.01, *<0.05)

Article Snippet: The published protocol from Takara’s SMART-Seq mRNA Single Cell LP User Manual was followed starting from Step V, First-Strand cDNA Synthesis.

Techniques: Functional Assay, Derivative Assay