Fig. S17 illustrates the biological differences between the various tumour cell subclusters in hepatocellular carcinoma. There is significant heterogeneity in the activity of VEGF, MAPK, EGFR, TGF-β, and hypoxia signalling across different tumour cell subclusters (Fig. S17C). Notably, the HCC_P02T, HCC_P01T and HCC_04T tumour cell subcluster exhibits heightened activity in the Wnt pathway, while the HCC_P08T tumour cell subpopulation shows increased activity in the TGF-β, EGFR, MAPK, and NFκB pathway, aligning with the results obtained during CytoTRACE analysis (Fig. S17D). The gene discussed is TGFB1; the disease is hepatocellular carcinoma.