Figure 8. Activation of AKT, AMPK and TOR pathways in normal and transformed cells.

(A) Time course expression and phosphorylation of the AKT protein in normal and transformed cell lines growing in media supplemented with different initial glutamine concentrations. For the protein expression analysis normal cells and transformed cells grown in media containing 4 mM glutamine and 0.5 mM glutamine, were collected at appropriate time points and 50 µg of proteins from the total cellular extract were subjected to SDS-PAGE followed by Western blotting with a specific anti-phosphorylated^Ser473 AKT (p-AKT) antibody and an anti-AKT (AKT) antibody. (B) Time course expression and phosphorylation of the AMPKα protein in normal and transformed cell lines growing in media supplemented with different initial glutamine concentrations. For the protein expression analysis normal cells and transformed cells grown in media containing 4 mM glutamine, 1 mM glutamine and 0.5 mM glutamine, were collected at appropriate time points and 50 µg of proteins from the total cellular extract were subjected to SDS-PAGE followed by Western blotting with a specific anti-phosphorylated^Thr172 AMPK (p-AMPK) antibody and an anti-AMPK (AMPK) antibody. (C) Time course expression and phosphorylation of the S6K protein in normal and transformed cell lines grown as described in B. Western blotting analysis has been performed by using a specific anti-phosphorylated^Thr389 S6K (p-AKT) antibody and an anti-S6K (S6K) antibody. The panels are representative of a triplicate analysis (Akt and AMPK) or duplicate analysis (S6K).