Fig. 6.

Therapeutic responses in a mouse orthotopic xenograft model. (A, B) Tumor volume was measured by bioluminescence imaging. Signal intensity was quantified as total photon flux from tissues. One-way ANOVA with Tukey’s post hoc test (*P < 0.05). (C) Survival probability for each group was estimated based on Kaplan–Meier curves. Log-rank test (P < 0.001); asterisks indicate groups with significant survival benefits based on Bonferroni-adjusted multiple comparisons (*P < 0.05). (D, E) Sections of the brains obtained from euthanized mice were immunostained for Zeb1 (pink) to identify invading cells (D). The number of infiltrated Zeb1+ cells (left side of yellow line in E), determined from 10 images captured for each mouse, was counted for each group (means ± SEM; ***P < 0.001 compared with controls; 2-way ANOVA with Tukey’s post hoc test). Expression levels of CD133, Sox2, N-cadherin, and β-catenin proteins were measured by immunohistochemistry (brown). For all images, hematoxylin (blue) was used to counterstain nuclei (yellow scale bar = 20 μm; black scale bar = 50 μm). (F) Schematic summary of study results. ATP generation by GBM TSs is suppressed by combined treatment with gossypol and phenformin, which target ALDH and OxPhos, respectively. This dual inhibition of tumor bioenergetics attenuates stemness, mesenchymal transition, and invasion, ultimately triggering cell death. Therapeutic responses were confirmed in a preclinical mouse model.