Multiple means of TOR pathway inhibition extend life span. (A) Removal of preferred amino acids from the media extends life span in proportion to the nutrient value of the eliminated amino acid, even though total nitrogen content is held constant. Removal of the preferred amino acid asparagine (integral = 3.24) or of the intermediately favored glutamate (integral = 2.56) confers increased survival compared with cells grown in synthetic complete media (integral = 1.99) (P < 0.001 for each amino acid drop out compared with synthetic complete). (B) Direct inhibition of the TOR pathway by low doses of rapamycin extends life span in a dose-responsive manner. The integrals under the life span curves are drug vehicle = 2.93, 100 pg/mL = 3.01, 300 pg/mL = 4.23, 600 pg/mL = 4.49, and 1 ng/mL = 4.51 (P < 0.001 for each dose >100 pg/mL vs. drug vehicle). (C) Inhibition of glutamine synthetase by the drug MSX extends life span by reducing intracellular glutamine, which reduces TOR signaling. The integrals under the life span curves are drug vehicle = 2.88, 30 μM = 2.95, 100 μM = 3.57, 200 μM = 4.11, 300 μM = 4.26, and 400 μM = 4.53 (P < 0.001 for each dose >30μM vs. drug vehicle).