Figure 5
From: HIF-independent role of prolyl hydroxylases in the cellular response to amino acids
Inhibition of PHDs blocks the response of mTORC1 to amino acids. (a, b) Phosphorylation state of S6K, S6 and 4EBP1 in cells starved for 2 h and then re-fed with amino acids for one more hour in the presence or absence of DMOG, rapamycin (Rap) or cobalt chloride (CC). (c) U2OS cells were transfected with a plasmid encoding control, nontargeting shRNA (shCntr) or cotransfected with three different plasmids encoding shRNAs for each member of the PHD family (shPHDs). After 48 h, cells were starved for amino acids for 2 h and then incubated for 20 min with (+ AA) or without (− AA) amino acids. The phosphorylation state of S6 was analyzed by western blot. (d) Cells were transfected as in (c) and starved for amino acids for 2 h and then incubated for the indicated time with amino acids (+ AA). The phosphorylation state of S6K was analyzed by western blot.
