Research Paper Volume 6, Issue 1 pp 48—57

Figure 6. Deregulated mTOR signaling contributes to accelerated aging of Bmal1^−/− mice. (a) Treatment with rapamycin suppresses TORC1 activity in both wild type and Bmal1^−/− (KO) cells. Cells were treated with indicated concentrations of rapamycin for 4 hrs, protein phosphorylation in cellular extracts were analyzed by western blotting procedure with antibodies recognizing the indicated proteins or protein modifications. U/t untreated cells. (b) Effect of rapamycin treatment on proliferation of wild type (black diamonds) and Bmal1^−/− (KO) (grey squares) cells. Cells were grown for 72 hrs in regular growth media with indicated concentrations of rapamycin. Cell biomass was assayed by crystal violet incorporation. Data represent average and standard deviations for 4 replicates. The experiment was repeated 3 times with similar results. a.u. – arbitrary units. * - statistically significant difference between genotypes. (c) Kaplan-Meyer survival curves of Bmal1^−/− mice treated with water (N = 73) or Rapatar in drinking water (N = 31). The difference between the survival curves is statistically significant according to logrank test.