Fig. 4

Reversible phenotypic changes. a Examples of adherent GBM cells in stem cell (Undiff_N) or differentiation (Diff_N) conditions in normoxia. See Supplementary Fig. 6A for hypoxia (scale bar = 200 µm). b Flow cytometric analysis of intracellular markers in control 3D sphere cultures (CTR), differentiation (Diff) conditions. N normoxia, H hypoxia. Black lines discriminate between negative and positive cells (mean + /− SEM, n = 3). Dotted line indicates mode expression in control cells. See also Supplementary Fig. 6B, C. c Distribution of subpopulations under different environmental conditions, phenotyping performed after 14 days of change (left) and 14 days after reverting to control culture (right). See Supplementary Data 1D for statistics and Supplementary Fig. 6D for more examples. d Distribution of subpopulations in xenografted NCH644 tumors in vivo (X) and after regrowth in vitro (De-X). Normoxia cultures are shown as control (CTR). See Supplementary Data 1F for statistics and Supplementary Fig. 6E, F for more examples. e Flow cytometric analysis of intracellular markers. Black lines discriminate between negative and positive cells (mean + /− SEM, n = 3). Dotted line indicate mode expression in control cells. f Kaplan–Meier survival curves of xenotransplanted mice. Subpopulations P2, P6, P11, and P15 were implanted directly after FACS. FACS-sorted bulk cells were used as control (CTR) (*p-value ≤ 0.05; **p-value ≤ 0.01, long-rank test). g Distribution of subpopulations in xenografted tumors. For each subpopulation, day of implant and day of mouse sacrifice are presented. See Supplementary Data 1H for statistics