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. Author manuscript; available in PMC: 2014 Nov 21.
Published in final edited form as: Mol Cell. 2013 Oct 10;52(4):574–582. doi: 10.1016/j.molcel.2013.09.018

Figure 2. Phase-dependent translational control of key cell cycle regulators including RICTOR and mTOR signaling.

Figure 2

(A) A heat map of the significance of translational regulation among cell cycle progression genes (asterisk: RICTOR). Shading represents significance level of increased or decreased translational regulation (red and blue, as indicated). (B) A diagram describing the 5′ UTR luciferase reporter assay (top panel). 5′ UTRs cloned into the firefly reporter are scaled to length. Capped mRNAs are transfected into synchronized cells prior to assaying reporter levels. Levels of translation directed by RICTOR or NUAK2 5′UTRs are shown: Y-axis is reporter value relative to cells in G1 (bottom panel). (C) Western blots showing RICTOR protein levels and phosphorylation of mTORC2 targets (AKT-S473, PKCα-S657). pHistone H3 is a mitosis marker. Tubulin is a loading control. (D) Mitotic progression of thymidine-synchronized MEFs: Y-axis is percent mitotic cells. Bars represent the mean +/- SD. See also Figure S2.