Research Paper Volume 12, Issue 1 pp 224—241

Figure 7. Downregulation of ASPM could arrest the cell cycle of GBM cells at the G0/G1 phase. (A, B) The enriched signalling pathways involved in ASPM-related genes by searching the LinkedOmics database. Cell cycle is the most enriched pathway. Volcano plot (A) and GSEA enrichment analysis (B) are shown. (C) Spearman’s correlation of the expression of cyclin E and ASPM in TCGA GBM, indicating a strong correlation. r = 0.53. (D–F) The cell cycle distribution of U87, U251 and U118 cells with stable ASPM knockdown was determined by PI staining and flow cytometry. Data shown are the mean ± SD (n = 3). G0/G1 phase distribution: *p < 0.05 versus control. (G, H) Western blot analysis of cyclin E and β-catenin protein levels in U87 and U118 cells with stable ASPM knockdown. (I) ASPM was immunoprecipitated with anti-cyclin E antibody from U87 cells, and the immunoprecipitates were subjected to Western blot analysis. (J) Luciferase activity of TOPFlash/FOPFlash in U87 cells that were transiently transfected with ASPM sh-NC or sh-2.