Fig. 1: Blocking AURKA stalls the proliferation of GBM by reducing glycolysis in a c-Myc dependent manner.

a Volcano plot of reverse-phase-protein-array (RPPA) data shows a reduced expression of c-Myc (red dot) in SF188 GBM cells treated with 100 nM alisertib for 24 h (n = 3 independent samples). FC: fold change. b Protein capillary electrophoresis of SF188 and GBM22 cells treated with the indicated concentrations of alisertib for 24 h. Vinculin is used as a loading control. c SF188 was transfected with non-targeting (siNT) or AURKA specific siRNA (siAURKA) or transduced with shARKA (shRNA) or sgAURKA (sgRNA) and the whole-cell protein lysates were subjected to protein capillary electrophoresis. d SF188 and GBM22 cells were transfected with non-targeting (siNT) or two specific siRNAs targeting Myc, treated with increasing concentrations of alisertib for 72 h, and cellular viability was analyzed (n = 4 independent samples). IC₅₀ in µM range. e SF188 and GBM22 cells were infected with empty vector or c-Myc adenovirus and treated with increasing concentration of alisertib for 72 h, and cellular viability was analyzed (n = 4 independent samples). IC₅₀ in µM range. f SF188 and GBM22 cell lysates were immunoprecipitated with IgG or c-Myc antibody and analyzed by standard western blot for the indicated antibodies. Input: cell lysate loading control. IgG: negative control. g SF188 cells were treated with DMSO or alisertib in the presence or absence of 5 µM MG132 and the whole-cell lysates were subjected to protein capillary electrophoresis for the indicated proteins. For h, i SF188 cells were treated with DMSO or alisertib in the presence or absence of 10 µg/mL cycloheximide (CHX) and the whole-cell lysates were subjected to protein capillary electrophoresis. Quantification of c-Myc protein level is shown in (i) (n = 3 independent samples). For j, k protein capillary electrophoresis for the indicated proteins of SF188 and GBM22 cells treated with DMSO or alisertib for different time points. Quantification of protein level is shown in (k) (n = 2 independent samples). l SF188 cells were transfected with c-Myc-WT or c-Myc mutant (T58A), treated with the indicated concentrations of alisertib for 24 h, and analyzed by protein capillary electrophoresis for the indicated proteins. m GBM22-Myc-WT or GBM22-T58A-Myc cells were implanted in the right striatum of nude mice. Two groups were randomly assigned: vehicle and alisertib after seven days of the implantation. Mice were treated three times per week and animal survival is provided (Kaplan−Meier-curve): GBM22-T58A-Myc-vehicle: 34d, GBM22-T58A-Myc-alisertib: 27d; GBM22-Myc-OE-vehicle: 38d, GBM22-Myc-OE-alisertib: 50d. The log-rank test was used to assess statistical significance (n = 5 independent samples) (*p = 0.0127, n.s not significant). n SF188 and GBM22 cells were transfected with non-targeting siNT or siAURKA in the presence or absence of 2 µM CHIR-908014 (CHIR) for 24 h and were subjected to protein capillary electrophoresis for the indicated proteins. o SF188 and GBM22 cells were transfected with HA-EV, HA-Aurora A-WT, HA-Aurora A-D274N and were subjected to protein capillary electrophoresis for the indicated proteins. Statistical significance was assessed by two-tailed student’s t-test in (m). Data are shown as mean ± SD in (d, e, i, k). Source data are provided as a Source Data file.