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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 6. Rac1 Interacts with mTORC1 and mTORC2 Proteins.

Figure 6

(A) Fifty to sixty percent confluent 293T cells were treated with PolyEthylImine (PEI) as a control or transfected with HA-tagged Rac1 or Rac2 and the different Myc-tagged mTORC proteins using PEI, as indicated.

(B) 293T cells were treated with PEI (control) or transfected with HA-tagged WT Rac1, V12Rac1, or N17Rac1 mutants and with either mTOR, Ric tor, or Raptor as indicated. Cells were then lysed and subjected to HA IP.

(C) 293T cells were transfected with either mTOR, Rictor or Raptor. Cells were lysed and subjected to pull-down assays using GST-fusion proteins of WT and Rac1 mutants for 1 hr at 4°C. The protein complexes were washed and then analyzed by SDS-PAGE and western blot. Results are representative of three experiments.

(D) 293T cells were transfected with either mTOR, Rictor, or Raptor. Cells were lysed and subjected to pull-down assays using GST-fusion proteins of WT and Rac1 mutants for 1 hr at 4°C. The protein complexes were washed and then analyzed by SDS–PAGE and western blot. Results are representative of three experiments.