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. 2011 Jul 1;7(7):737–747. doi: 10.4161/auto.7.7.15491

Figure 2.

Figure 2

ULK1 induces phosphorylation of Raptor on multiple sites in 293 cells. (A) A schematic showing the structure of Raptor, including the Raptor N-terminal conserved (RNC) domains, 3 HEAT repeats and 7 C-terminal WD40 repeats. The location of the phosphorylation sites examined in this study is shown in the central region of the protein. (B) HEK293 cells were transfected with HA-Raptor and either wild-type (WT) or kinase dead (KD) V5-ULK1. Following an HA immunoprecipitation, samples were assessed for phosphorylation of Raptor using site-specific antibodies. Total Raptor and ULK1 are shown as controls. (C) HEK293 cells were transfected with Myc-mTOR, HA-Raptor and either wild-type (WT) or kinase dead (KD) V5-ULK1 and treated with 1 µM Ku-0063794 for 3 h prior to lysis, where indicated. Immunoprecipitations and western blots were performed as in (B) with total mTOR, Raptor and ULK1 shown as controls.