Fig. 1

A IC₅₀ curves for dTRIM24 (MedChemExpress, USA) and IACS-9571 (MCE, hydrochloride form) in GBM#P3, GBM#BG7, GBM#06, and GBM#BG5 cells using the Cell Titer-Glo viability assay. B Quantification of tumoursphere formation assays for GSCs treated with different concentrations of dTRIM24 (0–10 μM) (A) or IACS-9571 (0—20 μM) for 6 days. GSCs (1000 cells/mL/well) were seeded in 6-well ultra-low adhesion plates. Inverted phase-contrast microscopy was used to count the sphere number. C Representative images of immunofluorescence staining for SOX2 (red; dilution 1: 100) or Nestin (green; dilution 1: 200) in GBM#P3 treated with dTRIM24 or IACS-9571 for 48 h. Nuclei were counterstained with DAPI (blue). Scale bar = 100 μm. D Western blot analyses of the TRIM24, SOX2, and GAPDH in lysates (20 µg) from GSCs treated with different concentrations dTRIM24 (0–10 μM) or IACS-9571 (0–20 μM). E Representative images of spheroids in 3D invasion assays for GBM#P3 GSCs treated with DMSO, dTRIM24 (5 μM), or IACS-9571 (10 μM), and evaluated at 24 h. Scale bar = 200 μm (lower). Graphic representation of the quantification of the distance of invading cells from the tumorspheres determined after 24 h (upper). F Relative cell viability for rescue experiments using the Cell Titer-Glo viability assay in GBM#P3 cells as indicated. Data are shown as mean ± SEM. Statistical significance was determined by one‐way ANOVA. *P < 0.05, **P < 0.01, ***P < 0.001