Skip to main content
. 2018 May 11;14(5):e1007369. doi: 10.1371/journal.pgen.1007369

Fig 1. Genetic manipulation of mTOR pathway alters circadian clock function.

Fig 1

RNAi knockdown of mTor lengthens the period length of circadian bioluminescence rhythms in MMH-D3 hepatocytes. (A) and 3T3-L1 adipocytes (B). Hepatocytes and adipocytes harboring the Per2-dLuc reporter were infected with lentiviral non-specific (NS) shRNA or shRNA constructs against mTor. Left panel: real-time bioluminescence expression was recorded in a Lumicycle luminometer on 35-mm culture dishes and the bioluminescence data are representative of at least three independent experiments. Middle panel: mTor knockdown efficiency was determined by Western blot analysis (middle). Right panel: period length and rhythm amplitude are mean ± standard deviation (SD) (n = 3 independent dishes). * p < 0.05 vs. NS. (C) Elevated mTOR via constitutively active Rheb shortens circadian bioluminescence rhythms in MMH-D3 hepatocytes. Bioluminescence rhythms from MMH-D3 hepatocytes harboring the Per2-dLuc reporter and overexpressing either the empty vector or constitutively active Rheb (CA-Rheb). p-S6 as a proxy of mTOR activation was hyper-phosphorylated in cells expressing CA-Rheb relative to vector control cells, whereas S6 levels were similar in the two cell lines. * p < 0.05 vs. Vector; ** p < 0.01 vs. Vector.