Fig. 4: Glucose deprivation (GD) induces tumor necroptosis in vitro and in vivo.

a Primary MVT-1 tumor cells were isolated at 5 weeks post-implantation and treated for 36 h with 0.5 mM glucose (GD) or 0.1 mM glutamine (GlnD) or 0.1% O₂ (hypoxia) and cell death was determined by PI staining using flow cytometry (left panel, n = 4 biologically independent samples, each) or representative of three independent experiments analyzed by western blotting using the indicated antibodies (right panel). Data are presented as mean values ± SEM. b Representative of three independent western blotting analysis. MVT-1 CRISPR CT or MVT-1 ZBP1 KO cells were treated with 5-Aza-2′-deoxycytidine (5-AD), for 3 days, followed by GD condition for 24 h (left panel) and determined by using the indicated antibodies. Cell death analysis of the cells treated with 5-AD for 3 days, followed by GD condition for 30 h, was determined by PI staining using flow cytometry (right panel, n = 4 biologically independent samples, each). Data are presented as mean values ± SEM. c Representative of three independent western blotting analysis. MVT-1 CRISPR CT or MVT-1 RIPK1 KO cells were treated with 5-AD for 3 days, followed by GD condition for the indicated time points and determined by using the indicated antibodies. d Representative of three independent western blotting analysis. MVT-1 cells were treated with 5-AD for 3 days, followed by treatment with 2-Deoxy-D-glucose (2DG) and determined by using the indicated antibodies. e FVB/NJ mice at three weeks post-implantation with MVT-1 cells were treated with vehicle or 2DG (i.p.) daily until week 5. Representative images of H&E and immunohistological stained sections with phospho-MLKL (p-MLKL) antibody. Scale bar, 50 µm. Two-sided student’s t test was used to determine the statistical significance of differences between groups. Differences with P values < 0.05 were considered significant. Source data are provided as a Source Data file.