Fig. 3

Glycolysis inhibition selectively impairs the growth of stem cell-like tumor subpopulations through induction of senescence but does not induce apoptosis or hamper stemness capacity. a CD133/PROM1^LOW and CD133/PROM1^HIGH patient-derived GBM cells were treated with increasing doses of 2-deoxyglucose (2-DG) from 0.25 to 3 mM. 24 h post-treatment cells were counted using trypan blue exclusion and then plotted as a percent of non-treated control cells. Statistical analysis was performed using two-sided students t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. b Western blot analysis of CD133/PROM1^HIGH patient-derived GBM cells following treatment with 1 mM of 2-DG for CASP3 (Pro-CASP3 35 kDa and cleaved CASP-3 17 & 19 kDa) and cleaved PARP (89 kDa). Positive control of cells treated with cytotoxic chemical were run in parallel. Graph represents densitometry quantification of western blots from three independent experiments. Statistical analysis was performed using two-sided students t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. c Western blot analysis of CD133/PROM1^HIGH patient-derived GBM cells following treatment with 1 mM of 2-DG for p21/CDKN1A expression. Graph represents densitometry quantification of western blots from three independent experiments. Statistical analysis was performed using two-sided students t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. d Non-treated controls and 2-DG (1 mM) treated CD133/PROM1^HIGH patient-derived GBM cells were subjected to β-galactosidase/GLB1 staining 24-h post-treatment to detect the presence of senescent cells. e Non-treated controls and 2-DG (1 mM) treated CD133/PROM1^HIGH patient-derived GBM cells were subjected to western blot analysis for the expression of BMI1. Graph represents densitometry quantification from three independent experiments. Statistical analysis was performed using two-sided students t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. f Non-treated controls and 2-DG (1 mM) treated CD133/PROM1^HIGH patient-derived GBM cells were subjected to neurosphere formation analysis and the graph represents the quantification of neurospheres > 50 µm in size. Statistical analysis was performed using two-sided students t-test. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant.