Fig. 4. Pib2 directly activates TORC1 in vitro in response to glutamine.

In vitro kinase assays were performed using TORC1 purified from Kog1-Tap expressing cells and bacterially-expressed GST-Pib2(304-635) with or without 30 mM L-glutamine. Dot plots represent the ratio of phosphorylated/total 4EBP1 normalized to sample with GST but without glutamine. Error bars represent the standard deviation (n = 4 independent experiments), and significance was determined by a two-tailed Student’s t-test.