Fig. 3: Colony formation and tumor-spheroid invasion assay.
A, B The clonogenic potential of 2D-cultured GBM cells was examined by seeding cells at a density of 1.0 × 103 per well. Thereafter, cells were treated for a total of 2 × 72 h and cultured for additional 14 days without treatment (=recovery). Biomass was quantified upon staining with 0.2% crystal violet and images were taken with a Leica DMI 4000B. A Representative light microscopic images. B Quantification of biomass (=cell colonies) using ImageJ software (colony count of treated cells; control >100 colonies/well, not presented). n = 3 independent experiments; *p < 0.0001; $p < 0.05; $$p < 0.01. One-way ANOVA. C Treatment schedule for the 3D invasion assay. D, E Analysis of invasive capacity into a matrigel-matrix. 3D-cultured GBM cells were treated with indicated substances or left untreated. GBM cells were monitored for a total of 15 days. Images were taken at a 5-day interval using a Leica DMI 4000B microscope (scale bar A, B: 50 μm). D Representative images of HROG05 Glioma spheres (left side) and Glioma stem-like cells (right side). E Invasive capacity (area of the spheres [µm2]) was examined with the oval selection function of FIJI-ImageJ according to [64]. *p < 0.05, **p < 0.01, ****p < 0.0001; $$$$p < 0.0001; §§§§p < 0.0001 dinaciclib; $$$$p < 0.0001. Two-way ANOVA.
