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. Author manuscript; available in PMC: 2011 Jan 21.
Published in final edited form as: Cell. 2010 Apr 8;141(2):290–303. doi: 10.1016/j.cell.2010.02.024

Figure 1. mTORC1 localizes to lysosomal membranes in an amino acid-dependent fashion while the Rag GTPases are constitutively localized to the same compartment.

Figure 1

(A) Images of HEK-293T cells co-immunostained for lysosomal protein LAMP2 (green) and mTOR (red). Cells were starved of and restimulated with amino acids for the indicated times before processing and imaging.

(B) Images of HEK-293T cells co-immunostained for LAMP2 (green) and raptor (red) Cells were treated and processed as in (A).

(C) Images of HEK-293T cells co-immunostained for LAMP2 (green) and RagC (red). Cells were treated and processed as in (A).

(D) RagC interacts with RagA and RagB independently of amino acid availability. RagC-immunoprecipitates were prepared from HEK-293T cells starved or stimulated with amino acids as in (A), and immunoprecipitates and lysates were analyzed by immunoblotting for the indicated proteins.

(E) Images of HEK-293T cells co-immunostained for RagA/B (green) and LAMP2 (red). Cells were treated, processed, and imaged as in (A).

(F) GFP-RagB and GFP-RagBGTP co-localize with co-expressed LAMP1-mRFP independently of amino acid availability. HEK-293T cells transfected with the indicated cDNAs were treated and processed as in (A).

(G) GFP-RagD and GFP-RagDGTP co-localize with co-expressed LAMP1-mRFP independently of amino acid availability. HEK-293T cells transfected with the indicated cDNAs were treated and processed as in (A). In all images, insets show selected fields that were magnified five times and their overlays. Scale bar is 10 μm.

See also Fig S1.