Fig. 4
5ALA + cells represent CD44 expressing mesenchymal cells. mRNA-based stemness index (mRNAsi) values across distinct GBM regions (Core, Rim, Invasive margin, 5ALA − and 5ALA +) for each patient are represented in a heatmap (A). Row (samples) and columns (GBM regions) are clustered by using a correlation algorithm. Comparison of the mRNAsi values according to brain regions (left) and TCGA-GBM samples (right) are shown as bar diagrams (B). The TCGA-GBM samples were pre-stratified according to GBM subtypes as described by Verhaak et al. [10]. Kruskal–Wallis tests showed a significantly higher mRNAsi in 5ALA + cells compared to Core and Rim, with p-values shown. Pearson correlation coefficient values between the mRNAsi and mRNA expression of selected genes are shown (C). The genes that showed a significant (p-value < 0.05) positive correlation with mRNAsi in 5ALA + cells for each patient were selected. tSNE clustering plots based on scRNA-seq dataset (Couturier et al.) are shown (D–G). The color code represents the single-cell wise UCell scores for different cell-signatures—glial progenitor cells (GP) (D), oligo-lineage cells (OLC) (E), mixed population including truncated radial glial cells, and cancer mesenchymal cells (F), and 5ALA + cells (G). tSNE clustering plots (Couturier et al.) with the color code representing the single-cell wise gene expression values for different marker genes—CD44 (H), AQP4 (I), FAM107A (J), and SOX9 (K), GLI3 (L), and TIMP1 (M) are shown. Heatmap showing the z-scored log2 TPM expression of selected marker genes (GIL3, TIMP1, FAM107A, SOX9, AQP4, and CD44) based on scRNA-seq dataset (Couturier et al.) across different cell types (5ALA + , GP, OLC, and mixed population) (N). qPCR validation results showing Log2 gene relative gene expression of CD44 gene in spatially resolved RNA profiles across Core, Invasive margin, and 5ALA + cells (O). P-values are shown as calculated by paired T-test