Figure 4
Acute glutamine depletion and ammonia regulate AMPK and mTORC1 signalling. Cells (HEK293, Huh7, T47D and MCF7) were plated in complete medium containing glutamine for 24 h. One culture was lysed and collected for western blotting 24 h later (T0). The remainder of the cultures were washed with DPBS and fresh glutamine-depleted media with or without 0.8 mM NH4Cl was added. Cultures were collected and lysed at 15 min for western blotting. Levels of phosphorylated S6K (T389) relative to total S6K protein levels (A) and phosphorylated ACC (S79) relative to total ACC protein levels (B) were quantified using western blot. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001.
