Fig. 3: Ferroptosis is a metabolic vulnerability of differentiated glioma cells (DGC). | Nature Communications

Fig. 3: Ferroptosis is a metabolic vulnerability of differentiated glioma cells (DGC).

From: Therapeutic targeting of differentiation-state dependent metabolic vulnerabilities in diffuse midline glioma

Fig. 3

A Differential expression of epithelial-mesenchymal transition (EMT) genes by bulk transcriptomics in DIPG-007, SF7761, and DIPG-XIII gliomaspheres (GS) vs. DGC. Data presented from three biological replicates. B Simplified scheme depicting the role of glutathione peroxidase 4 (GPX4) in ferroptosis. C, D RSL3 dose-response in (C) DGC and (D) GS DIPG-007, SF7761, and DIPG-XIII with or without 1 h ferrostatin-1 (Fer-1) pre-treatment. E Flow cytometry assessment of intracellular lipid reactive oxygen species (ROS) using C-11 BODIPY in DIPG-007 DGC treated with RSL3 for 6 h with or without 1 h Fer-1 pre-treatment. Data expressed as mean ± SD mean fluorescent intensity (MFI) (****p < 0.0001; multiple unpaired t test). F Cell viability of RSL3-treated DIPG-007 DGC in the presence of bafilomycin A1 (Baf-1), necrosulfonamide (NSA), ZVAD-FMK (Z-VAD), Fer-1, or Trolox. G RSL3 dose response in DIPG-007 GS cultured in antioxidant-free B-27-supplemented GS, DGC, or reduced-serum (2.5%) DGC media, with or without 1 h Fer-1 pre-treatment. Cell viability (C, D, G) assessed with Cell Titer-Glo 2.0 (DGC) or Cell Titer-Glo 3D (GS) at 48 h and data expressed as percent vehicle control (0.1% DMSO). Error bars represent mean ± SD from three biological replicates with ns = not significant, *p < 0.05, **p < 0.01, ****p < 0.0001 by area under proliferation curve test followed by either two-tailed Student’s t-test) (C, D) or one-way ANOVA for multiple Uncorrected Fisher’s LSD test (G). For panels E and F, error bars represent mean ± SD from three biological replicates with nd = no discovery, *p < 0.00001 by multiple unpaired t-test and Two-stage step-up. Panels C–G are representative of data from three biological replicates. TGFB2, transforming growth factor beta-2; VCAM1, vascular cell adhesion molecule 1; TAGLN, transgelin; ACTA2, actin alpha 2; COL11A1, collagen type XI alpha 1 chain; SNAI1, snail family transcriptional repressor 1; MMP2, matrix metallopeptidase 2; MMP11, matrix metallopeptidase 11; GSH, glutathione. For panels A and C-F, red indicates GS; blue, DGC. Figure 3B created with BioRender.com released under a Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International license.

Back to article page