Fig. 4
Glycolysis inhibition selectively induces autophagy in CD133/PROM1HIGH stem cell-like patient-derived GBM cells but not CD133/PROM1LOW non-stem-like patient-derived GBM cells. a Schematic diagram depicting the mechanism of action for chloroquine (CQ) and the principal of the autophagy flux assay. Created using Biorender.com. b CD133/PROM1HIGH patient-derived GBM cells were treated with 2-DG (1 mM) and subjected to autophagy flux analysis where cells were treated with the late-stage autophagy inhibitor CQ and subjected to western blot analysis for the autophagosome-associated proteins SQSTM1, MAP1LC3A-II (14 kDa), and MAP1LC3B-II (14 kDa). Graph represents densitometry quantification of western blots from at least 3 independent experiments. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. c CD133/PROM1HIGH patient-derived GBM cells with exogenous overexpression of EGFP-LC3 were treated with 2-DG (1 mM) and/or the late-stage autophagy inhibitor CQ and autophagosome assembly was assessed by monitoring the formation of EGFP-LC3+ punctae and the number of EGFP-LC3+ punctae were quantified. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. d CD133/PROM1LOW patient-derived GBM cells were treated with 2-DG (1 mM) and subjected to autophagy flux analysis where cells were treated with the late-stage autophagy inhibitor CQ and subjected to western blot analysis for the autophagosome-associated proteins SQSTM1, MAP1LC3A-II (14 kDa), and MAP1LC3B-II (14 kDa). Graph represents densitometry quantification of western blots from at least 3 independent experiments. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant. e CD133/PROM1LOW patient-derived GBM cells with exogenous overexpression of EGFP-LC3 were treated with 2-DG (1 mM) and/or the late-stage autophagy inhibitor CQ and autophagosome assembly was assessed by monitoring the formation of EGFP-LC3+ punctae and the number of EGFP-LC3+ punctae were quantified. Statistical analysis was performed using ANOVA. *p < 0.05; **p < 0.01; ***p < 0.001; ns = non-significant