Fig. 3

Adaptation of GBM subpopulations to hypoxia. a Distribution of subpopulations in hypoxia (H). Normoxia (N) is shown as control (CTR) and hypoxia (H) at 16 h, 48 h, 7 days, and 60 days. Data shown for NCH644, additional cultures in Supplementary Fig. 4C. See statistics in Supplementary Data 1B, color code in Fig. 1e. b Self-renewal test of 16 subpopulations in hypoxia, including sphere formation (mean + /− SEM) and sphere diameter (Box limits indicate the 25th and 75th percentiles and center lines show the medians as determined by R software; whiskers represent the extreme low and high observed values, unless those are above 1.5 times interquartile range (IQR)—thereby whiskers are limited to 1.5 IQR. All outlying data points are represented by dots). Bulk cells were used as control (CTR). Statistical differences within the same subpopulations are shown (*p-value ≤ 0.05, Kruskal–Wallis test, see Supplementary Data 4B for statistics). c Proliferation test in hypoxia (mean doubling time + /− SEM). Bulk cells were used as control (CTR). Statistical difference was only found between P7 and P13 (mixed linear model). d Distribution of 16 subpopulations at day 0 (left) and after 60 days in hypoxia (right) (see Supplementary Data 2B for statistics). e Markov modeling of state transitions in hypoxia. Arrows represent direct state transitions between subpopulations, thickness of lines corresponds to transition probabilities. See Supplementary Fig. 5E for transition matrix. f Predicted time needed to reach equilibrium for each subpopulation in hypoxia