Fig. 3: Correlation of cancer stem cells (CSC) with worse prognosis and undesirable immunotherapy response.

a Scatter plots showing the correlation between the identified CSC signature and CytoTRACE score in the discovery dataset and two validation datasets (GSE166635, GSE146115). b Distribution of CSC in normal and HCC tissues. c Distribution of CSC in early and advanced stage HCC tissues. d Immunohistochemistry images of five proteins identified in the CSC signature (ENO1, TKT, CD63, G6PD, and KRT8) in our cohort consisting of 32 HCC patients. (e) Boxplots displaying the distribution of the five proteins in early and advanced stage samples. Statistical significance (p value): ‘*’ for ‘between 0.05 and 0.01’, ‘**’ for ‘between 0.01 and 0.001’ and ‘***’ for ‘lower than 0.001’. f HCC patients in the TCGA cohort with higher CSC infiltration exhibited shorter overall survival (OS). g CSC score showed positive correlation with Macrophage M2 and negative correlation with CD8 T cells. h The non-responders exhibited higher CSC scores compared to responders in HCC patients treated with immunotherapy. i Pathway enrichment analysis revealing that the signature of CSC was enriched in non-responders in multiple immunotherapy cohorts.