Fig. 2
Inhibition of AMPK sensitizes GB cells to nutrient starvation and hypoxia
(A) LNT-229, LN-308 and G55T2 wildtype (wt) and AMPK catalytic subunits double knockout (DKO) cells were treated in serum- and glucose-free DMEM for 24 h. Cell death was analyzed by PI staining and quantified by flow cytometry (n = 3, mean ± SD, **p < 0.01, Student’s t-test). (B) Cell death of LNT-229, LN-308 and G55T2 wildtype and AMPK DKO was analyzed by an LDH release assay after incubation of the cells in serum-free DMEM containing 2 mM glucose in normoxia or hypoxia (0.1% O2) (n = 4, mean ± SD, *p < 0.05, **p < 0.01, Student’s t-test). (C) Primary GB cells (P3NS) were treated with vehicle (DMSO), 1 µM BAY974 or 1 µM BAY3827 in serum-free medium containing 2 mM glucose for 8 h in normoxia or hypoxia (0.1% O2). Cellular lysates were analyzed by immunoblot with antibodies for P-ACC, P-AMPK, AMPKα1/2 and actin. (D) Human primary glioblastoma cells P3NS, NCH60 and NCH644 cells were treated with vehicle (DMSO), 1 µM BAY974 or 1 µM BAY3827 in serum-free medium without glucose. Cell death was analyzed by PI staining and flow cytometry (n = 3, mean ± SD, **p < 0.01, Student’s t-test)