Molecular characteristics of MBL and derived stem like cells. (A) Heat map showing mRNA levels of the indicated genes in primary MBL (pMBL), scalp (M1) and neck (M2) metastases compared to normal cerebella (average of n = 8) as control (CTRLs). Genes are grouped depending on the molecular subgroups, which they identify (SHH, WNT, GROUP-3, GROUP-4). A green-red colour scale depicts normalized Delta Ct values (green, lower expression, red, higher expression). (B-C) Flow cytometry analysis (FACS) of CD133 in the starting population from neck metastasis (hMED bulk) (B) and after 14 days of culture (C) (isotypic control not shown). P3 shows percentage of positive cells while P4 shows percentage of negative cells. (D) Representative bright field image of neurospheres derived from neck metastasis after 14 days of culture. (E) Histograms showing mRNA levels of CD133 in MBL-neck metastasis (bulk) and in its derived SLC compared to normal cerebella as control (CTRLs). (F) Immunofluorescence staining with anti-Nestin and anti-Sox2 (green), Hoechst staining (blue) and merge of SLC. Scale bar = 5 μm for all panels. (G) Heat maps of expression levels of the indicated genes belonging to the highlighted categories in: pMBL, metastases (M1 and M2), in SLC derived from M2 and in normal adult cerebella as control (CTRLs). A green-red colour scale depicts normalized Delta Ct values (green, lower expression, red, higher expression).