Extended Data Fig. 5: MK-8931 Treatment Shows Little Effect on Glioma Cells in Vitro.
a,b, Representative images (a) of tumorspheres derived from CCF-3264 GSCs treated with MK-8931 (50 μg/mL) or vehicle control. Quantifications (b) show the number and size of tumorspheres treated with MK-8931 or control. Data are shown as mean ± SEM. Statistical significance was determined by two-tailed Student’s t-test. n = 3 independent experiments; ns, not significant. c, Immunoblot analysis of SOX2 expression in GSCs (CCF-3264 and CCF-DI315) treated with MK-8931 (50 μg/mL) or the vehicle control for three days. GAPDH was blotted as control. Similar results were confirmed in 3 independent experiments. d, Immunoblot analyses of cleaved PARP and cleaved caspase 3 to detect apoptosis in GSCs (CCF-3264 and CCF-DI315) treated with MK-8931 (50 μg/mL), etoposide (Etop, 1 μM), or the vehicle control for indicated times. GAPDH was blotted as control. Ctl: Control; Etop: Etoposide. Similar results were confirmed in 3 independent experiments. e, Cell viability assay of GSCs (CCF-3264 and CCF-DI315) treated with MK-8931 (10 or 50 μg/mL) or the vehicle control for three days to examine the effect of MK-8931 treatment on glioma cells. Data are shown as mean ± SEM. Significance was determined by one-way ANOVA analysis. n = 4 independent experiments; ns, not significant. f,g, RNA-seq analyses of GSCs treated with MK-8931 (50 μg/mL) or the vehicle control. The volcano plot (f) shows differentially and non-differentially expressed genes. Green dots represent the up-regulated genes (40); red dots represent the down-regulated genes (57); and blue dots represent the unaffected genes (24289) by MK-8931 treatment in GSCs. Gene ontology analysis (g) predicts differentially expressed genes enriched in the indicated cellular processes, but none of them was significantly affected by MK-8931 treatment in GSCs.
