Fig. 2.

Glioblastoma invasion depends on PFKM. (A, B) Exposure (24 hours) of high glucose (HG: 5 µg/mL) but not low glucose (LG: 1 µg/mL) induces PFKM protein and activity in GBM cells. Loading control: GAPDH. (C) HG treatment (HG: 5 µg/mL) increases migratory capacity of GBM cells exposed to bevacizumab (BEV: 50 µg/mL) in a wound healing assay. LG (1 µg/mL). Data shown are normalized to 0 hours. Scale bar = 150 µm. (D) PFKM-silencing inhibits GBM cell migration in wound healing assay. (E) PFKM-silencing impairs GBM cell invasion in a 3D spheroid assay. Data shown are normalized to 0 hours. Scale bar = 100 µm. (F) PFKM-silencing (shPFKM#1 & shPFKM#2 vs shCONT) extends the survival for tumor-bearing mice (n = 5 per arm). (G) Immunoblot analysis of PFKM protein level in PFKM^high and PFKM^low T10 sub-cell lines used in (H). (H) Kaplan-Meier survival analysis of animals intracranially injected with PFKM^high or PFKM^low T10 cells and treated intraperitoneally with PBS or BEV (5 mg/kg). (I) Representative H&E staining of coronal section of brains from study shown in (H). Scale bar = 2.5 µm. (J) Bevacizumab (BEV: 5 mg/kg) treatment drives superior survival benefits in mice intracranially implanted with PFKM^low T10 cells.

Significance is determined by log-rank test (F, H) or t test (B–E), where data are shown as mean ± SEM (*P < .05, **P < .01, ***P < .001).