Figure 7
Chronic glutamine depletion and ammonia regulate AMPK and mTORC1 signalling in HEK293 and T47D cells. HEK293 and T47D cells were plated in complete medium containing glutamine for 24 h. Cultures were washed with DPBS and fresh glutamine-depleted media with or without NH4Cl was added. Cultures were collected 3 days later and protein was extracted for western blotting. Protein abundance of phosphorylated ACC (S79) or S6K (T389) relative to total ACC or S6K in HEK293 (A) and T47D (B) cells 3 days post-incubation was quantified using western blot. Blots shown were representative from 4 independent experiments. *p ≤ 0.05, **p ≤ 0.01, ***p ≤ 0.001. Full length blots are presented in Supplementary information Fig. 4.
