Fig. 5

NDRG1 downregulation in CSCs mirrors the ASCL1-induced phenotype, whereas its overexpression promotes PN-to-MES transition (PMT). a NDRG1 silencing in PN CSCs results in significant AKT hypoactivation (WB). b qPCR analysis of gene classifiers in NDRG1-silenced CSCs (red-lined and red-filled bars in the histograms) shows the promotion of the PN and the further reduction of the MES phenotypes, as observed upon ASCL1 overexpression (black-lined and black-filled bars in the histograms). c Intracranial xenografts, induced by NDRG1-silenced CSCs and analyzed 3–4 months after transplantation, are significantly smaller than controls (L0605 and L0512; human-specific EGFR staining: brown, ×20). d NDRG1-silenced CSC-derived xenografts are characterized by the enhancement of PN features, such as the presence of focal areas containing neuronal-like cells organized as rosettes (H&E, ×400), and the increased immunoreactivity for the PN markers Olig2 and PDGFRα (all markers stained in brown, ×400; inset, ×800). e Upregulation of MES markers and reduction in PN ones are observed upon NDRG1 transduction of GBM CSCs (qPCR). f NDRG1-overexpressing tumors display increased MES morphological features, such as the development of areas made up by spindle-shaped cells with large and elongated nuclei (H&E, ×400; inset, ×800), as well as increased MES marker immunoreactivity (all markers stained in brown, ×400)