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. Author manuscript; available in PMC: 2011 Jun 11.
Published in final edited form as: Mol Cell. 2010 Jun 11;38(5):768–774. doi: 10.1016/j.molcel.2010.05.017

Figure 1.

Figure 1

mTORC1 purification. (A) mTORC1 and FLAG-raptor were purified by tandem gel filtration chromatography. Their masses were estimated based on known molecular weight standards as indicated (Thyroglobulin (669 kDa), Ferritin (440 kDa), Aldolase (158 kDa) and Conalbumin (75 kDa)). (B) The gel filtration fraction corresponding to mTORC1 was analyzed by SDS-PAGE followed by silver and Coomassie staining as well as immunoblotting for indicated proteins. (C) Schematics of mTORC1 components illustrating the various predicted domains. (D) In vitro kinase assay showing that purified mTORC1 phosphorylates S6K1 and is inhibited by both rapamycin-FKBP12 (rapa) and Torin1. (E) EM of negatively stained mTORC1. A raw image of mTORC1 particles (circled) and a representative class average from the classification of 10,080 particles (inset). The scale bar represents 50 nm, and the side length of the panel showing the class average is 45 nm. “see also Figure S1”