Fig. 6
Combination of glycolysis and autophagy inhibition cumulatively decreases the growth of stem cell-like tumor subpopulations and suppresses stemness by blocking induction of senescence and promoting apoptosis. a CD133/PROM1HIGH patient-derived GBM cells were treated with 2-DG (1 mM) and/or the autophagy inhibitor Spautin-1 (10 µM) for 24 h and the number of viable cells were counted using trypan blue exclusion. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001. b An additional independent CD133/PROM1HIGH patient-derived GBM cell line, GBM8, was treated with 2-DG (1 mM) and/or the autophagy inhibitor Spautin-1 (10 µM) for 24 h and the number of viable cells were counted using trypan blue exclusion. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001. c Normal human astrocytes were treated with a combination of 2-DG (1 mM) and the autophagy inhibitor Spautin-1 (10 µM) for 24 h and the number of viable cells were counted using trypan blue exclusion. Statistical analysis was performed using two-sided students t-test. *p < 0.05; **p < 0.01; ***p < 0.001. d CD133/PROM1HIGH patient-derived GBM cells were treated with 2-DG (1 mM) and/or Spautin-1 (10 µM) and subjected to western blot analysis for CASP3 (Pro-CASP 35 kDa and cleaved CASP-3 17 & 19 kDa), cleaved PARP (89 kDa), and p21/CDKN1A. Graph represents densitometry quantification from three independent experiments. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. e CD133/PROM1HIGH patient-derived GBM cells were treated with 2-DG (1 mM) and/or Spautin-1 (10 µM) and subjected to neurosphere formation analysis and the graph represents the quantification of neurospheres > 50 µm. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001 ns = non-significant. f CD133/PROM1HIGH patient-derived GBM cells were treated with 2-DG (1 mM) and/or Spautin-1 (10 µM) and subjected to western blot analysis for the expression of BMI1. Graph represents densitometry quantification from three independent experiments. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant