Fig. 2

SKF83566 inhibits GBM invasion in vitro and in vivo. (A) Representative images from co-culture invasion assays for P3, BG5, and BG7 human GSCs treated with SKF83566 (Scale bar = 100 μm) and (B) quantitative representation of invasive ratios. Data are shown as the mean ± SEM. Statistical significance was determined by the unpaired Student’s t-test. ***P < 0.001. (C and D) Representative images of spheroids in 3D invasion assays for P3, BG5, and BG7 human GSCs treated with SKF83566, and evaluated at 96 h. Scale bar = 200 μm. Graphic representation of invasive ratios from 3D invasion assays for P3, BG5 and BG7 human GSCs treated with SKF83566. The quantification of the distance of invading cells from the tumor spheres was determined after 96 h. Data are shown as the mean ± SEM. The unpaired Student’s t-test determined statistical significance. ***P < 0.001. (E) Schematic depiction of the schedule for implantation and drug treatment in vivo. Ten days after implantation of tumor cells, mice were treated with SKF83566 by intraperitoneal injection (20 mg/kg/day). Bioluminescence imaging (BLI) was performed at days 10, 17, 24 and 31. Nude mouse image was obtained from the Biorender webpage. (F and G) Bioluminescence images and the corresponding quantification of bioluminescence intensities, reflecting tumor burden, were obtained in mice implanted with P3 cells at days 10, 17 and 24 (n = 5 per group). Data are shown as mean ± SEM. Statistical significance was determined with ANOVA. ***P < 0.001. (H) The survival curves of tumor-bearing mice implanted with P3 cells treated with SKF83566 or vehicle control (DMSO) (n = 5 per group). A log-rank test was used to assess statistical significance. Data are shown as mean ± SEM. **P < 0.01. (I) Representative hematoxylin and eosin staining of mouse brains implanted with P3. (I above) Scale bar = 2.5 mm, (I below) Scale bar = 500 μm. (J) Quantification of immunohistochemical staining for Ki67 in sections from P3 and BG5 xenografts. Data are shown as the mean ± SEM. Unpaired Student’s t-test determined statistical significance. ***P < 0.001