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. Author manuscript; available in PMC: 2013 Jul 31.
Published in final edited form as: Science. 2013 May 31;340(6136):1100–1106. doi: 10.1126/science.1232044

Figure 2.

Figure 2

The GATOR complex is required for the regulation of the TORC1 pathway by amino acids. (A) shRNA-mediated depletion of the GATOR2 components Seh1L, WDR24, or WDR59 in HEK-293T cells inhibits amino acid-induced S6K1 phosphorylation. (B) In HEK-293Ts expressing shRNAs targeting the GATOR1 components DEPDC5, Nprl2, and Nprl3, S6K1 phosphorylation is insensitive to amino acid withdrawal. In (A) and (B) cells were starved of amino acids for 50 min or starved and restimulated with amino acids for 10 min. Cell lysates were immunoblotted for the phosphorylation state of S6K1. dsRNA-mediated depletion in S2 cells of (C) dSeh1L; (D) dWDR59 and dWDR24; and (E) dDEPDC5, dNprl2, and dNprl3. S2 cells were treated with the indicated dsRNAs and were starved of amino acids for 90 min or starved and restimulated with amino acids for 30 min. Immunoblotting was used to detect the phosphorylation state of dS6K. (F) S2 cell sizes after dsRNA-mediated depletion of dSeh1L and dDEPDC5. (G) GATOR2 functions upstream of GATOR1. S2 cells were treated with the indicated combinations of dsRNAs and starved and restimulated with amino acids and analyzed as in (C)–(E). (H) Schematic depicting the relationship between GATOR1 and GATOR2 in their regulation of mTORC1.