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. Author manuscript; available in PMC: 2013 Aug 23.
Published in final edited form as: Mol Cell. 2011 Apr 8;42(1):50–61. doi: 10.1016/j.molcel.2011.03.017

Figure 7. The C-Terminal Region of Rac1 Is Required for the Direct Interaction with mTOR.

Figure 7

(A) Fifty to sixty percent confluent 293T cells were treated with PEI (control) or transfected with mTOR, Rictor, or Raptor. Cells were then lysed and biotin-peptides of the Rac1, Rac2, and Cdc42 C-terminal domains and mutated Rac1 peptide: PPP/AAA or RKR/AAA was used to precipitate mTORC proteins for 1 hr at 4°C in the presence of streptavidin beads.

(B and C) 293T cells were transfected with Flag-mTOR. The Flag IPs were eluted by the Flag peptide. The free mTOR protein was precipitated by biotin-Rac1 C-terminal peptide or the biotin-RKR/AAA mutant, followed by streptavidin agarose. Samples were analyzed by SDS-PAGE and western blot (b) or the SDS-PAGE was silver stained (C). MW: molecular weight markers, WL: total proteins from Flag-mTOR transfected cells (1) WT biotin-Rac1 peptide; (2) biotin RKR/AAA mutant; (3) Flag IP. Data are representative of three experiments.