Fig. 8

Trehalose modulates mTOR, AMPK and ERK1/2 activity. U373-MG (A, C, E) and T98G cells (B, D, F) were untreated (C) or treated with 90 mM trehalose (TRE) for different experimental times. mTOR activity (A, B) was evaluated as ratio between p70 S6K1 phosphorylated on Thr389 residue (P-p70) and total p70 S6K1 (p70); AMPK activity (C, D) was evaluated as ratio between the catalytic subunit AMPKa phosphorylated on Thr172 residue (P-AMPKa) and total AMPKa (AMPKa); ERK 1/2 activation (E, F) was evaluated as ratio between ERK2 phosphorylated on Thr202/Tyr204 residues (P-ERK) and total ERK2 (ERK2). For all of these proteins, after detection of the phosphorylated form, membranes were stripped and reprobed for the expression of the total protein. The ratio phosphorilated protein/total protein of trehalose-treated cells was expressed as fold-change over the ratio of the control sample at the corresponding experimental time. Results are presented as mean ± SE from 3 to 5 independent experiments. *p < 0.05, and **p < 0.01, significantly different from control cells at the corresponding time. Typical western blots are shown