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. Author manuscript; available in PMC: 2010 Dec 7.
Published in final edited form as: Nat Genet. 2010 Feb 14;42(3):229–233. doi: 10.1038/ng.533

Fig.4. TMEM127 colocalizes with amino acid-activated mTORC1.

Fig.4

A) Effect of 1-hour amino acid (AA) starvation, followed or not by replenishment (15 min), on S6K phosphorylation of 293E cells with TMEM127 knockdown by two independent shRNA sequences, T1 and T2. A control knockdown (C) is shown for comparison. B) Effect of amino acid (AA) treatment as in "A" on 4EBP1 phosphorylation of 293E cells overexpressing HA-TMEM127 or empty vector (EV). HA indicates TMEM127 expression. β-actin is the loading control. P-S6K, phosphorylated S6 kinase at residue T389; P-4EBP1, phosphorylated 4EBP1 at residues T37-46; S6K, total S6 kinase. C) HEK293T cells co-transfected with HA-TMEM127 and myc-mTOR were serum-starved overnight, depleted of amino acids for 1h followed by reexposure for 15 minutes (AA+). Cells were fixed and stained with antibodies specific for HA (green), myc (red) or DAPI (blue) before imaging. myc-mTOR is diffusely present in the cytoplasm in the absence of amino acid and becomes localized to a perinuclear region where TMEM127 is detected upon amino acid exposure. Similar results were obtained with two independent TMEM127 constructs. Bars, 5μm.