Extended Data Fig. 6: The Raptor-S606D mutation affects its interaction with Rheb and DEPTOR in a LATS-dependent manner.
From: LATS suppresses mTORC1 activity to directly coordinate Hippo and mTORC1 pathways in growth control
a,b, The Raptor-S606D mutation does not affect mTORC1 complex formation. IB analysis of WCLs and IP derived from HEK293 CRISPR knock-in cells (a) or HEK293 cells transfected with indicated constructs (b). c, The S606D mutation impairs its interaction with Rheb. IB analysis of WCLs and IP derived from indicated CRISPR knock-in cells expressing HA-tag Rheb. d, The Raptor-S606D mutation does not affect its interaction with Rag GTPases. IB analysis of WCLs and IP derived from indicated HEK293 CRISPR knock-in cells. e, The Raptor-S606D mutation increases its interaction with DEPTOR. IB analysis of WCLs and IP derived from HEK293 cells transfected with indicated constructs. f, The Raptor-S606D mutation minimally affects its interaction with PRAS40. IB analysis of WCLs and IP derived from indicated HEK293 CRISPR knock-in cells. g, IB analysis of WCLs and IP derived from HEK293 cells transfected with indicated constructs. h, IB analysis of WCLs and GST pull-down products (GST-PD) derived from HEK293 cells transfected with indicated constructs. i,j, IB analysis of WCLs and IP derived from indicated HEK293 CRISPR knock-in cells stably expressing HA-Rap2A (as a negative control) or HA-Rheb. The cells were treated with 1 μg/ml LatB (i) or 10 μM FSK (j) for 60 min before harvesting. k, IB analysis of WCLs and IP derived from HEK293 cells stably expressing HA-Rheb or HA-DEPTOR cultured at low (30%) or high (100%) cell density. l,m, IB analysis of WCL and IP derived from indicated HEK293 CRISPR knock-in cells. The cells were treated with 1 μg/ml LatB (l) or 10 μM FSK (m) for 60 min before harvesting. Western blots in a–m were performed n = 2 independent experiments, with similar results obtained. Unprocessed immunoblots are shown in Source Data Extended Data Fig. 6.
