Fig. 3
Retransfection of the catalytic subunit of AMPK in knockout cells restores adaptation to starvation conditions
(A) LNT-229 wildtype (wt) and AMPK catalytic subunits double knockout (DKO) cells were stably transfected with empty vector (control) or PRKAA2. Cellular lysates were analyzed by immunoblot with antibodies for AMPK α1, AMPK α2 and actin. (B) Immunoblot analysis of LNT-229 wildtype and AMPK DKO PRKAA2 lysates treated in serum-free medium with 2 mM glucose for 8 h in normoxia or hypoxia (0.1% O2) was performed with antibodies for P-ACC, AMPK α2 and actin. (C) LNT-229 wildtype and AMPK DKO PRKAA2 cells were treated in serum-free medium without glucose. Cell death was analyzed by PI uptake and flow cytometry (n = 3, mean ± SD, n.s. not significant, **p < 0.01, Student’s t-test). (D) Cell death of LNT-229 wildtype and AMPK DKO cells was analyzed by LDH release assay after incubation in serum-free medium supplemented with 2 mM glucose in normoxia and hypoxia (0.1% O2) (n = 4, mean ± SD, *p < 0.05, **p < 0.01, Student’s t-test)