Research Paper Volume 6, Issue 1 pp 48—57

BMAL1-dependent regulation of the mTOR signaling pathway delays aging

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Figure 1. BMAL1 is negative regulator of TORC1 activity in cells. (a) Phosphorylation and total protein level of ribosomal S6 protein in primary lung fibroblasts isolated from wild type and Bmal1−/− mice. Protein phosphorylation in cellular extracts were assayed by western blotting procedure with antibodies recognizing the indicated proteins or protein modifications WT1, WT2, KO1, KO2 and KO3 represent independently isolated populations fibroblasts isolated from wild type (WT) and Bmal1−/− (KO) mice. Cells were incubated in DMEM with 10% FBS (+) or serum deprived for 24 hrs (−). (b) and (c) Wild type (WT) and Bmal1−/− (KO) fibroblasts were subject to amino acids (b) or serum (c) withdrawal for indicated period of time. Kinetics of changes in phosphorylation of mTORC1 downstream targets are shown on the representative western blotting.