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Cell. Author manuscript; available in PMC 2015 Mar 2.
Published in final edited form as:
Cell. 2014 Feb 13; 156(4): 786–799.
doi: 10.1016/j.cell.2014.01.024
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Figure 1

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Conserved binding of TSC2 to the Rag GTPase complex

(A) Drosophila Rag proteins are required for complete TORC1 inactivation upon amino acid withdrawal. Immunoblots of S2 cells treated with dsRNAs against LacZ (control), RagA or RagC, incubated with media lacking only amino acids in the presence of dialyzed serum for indicated times.

(B) Shot-gun mass spectroscopy analysis identifies Tsc2 as a Rag binding protein. Two statistics of peptide abundance are shown for Tsc2 and control proteins from two independent FLAG-RagA + FLAG-RagC immunoprecipitates (IP) from S2 cells.

(C) Confirmation by coIP that Rag proteins bind Tsc2 but not an unrelated protein, Medea. Immunoblots of anti-FLAG IPs from Drosophila S2 cells expressing FLAG-tagged RagA + RagC and V5-tagged Tsc2 or Medea, as indicated.

(D) Human TSC2 binds human Rag dimeric complexes. Immunoblots of anti-FLAG IPs from HEK293FT cells expressing FLAG-tagged TSC2 and HA-GST-tagged Rag proteins or an HA-GST control, as indicated.

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