Figure 3. TBK1 promotes mTORC1 signaling independently of mTORC2‐dependent Akt S473 phosphorylation.

1. mTORC1 signaling remains intact in MEFs lacking mTORC2 function. Rictor^−/− MEFs stably expressing vector control (V) or rescued stably with HA‐rictor were serum‐starved (20 h), pre‐treated with Ku‐0063794 [1 μM] (30 min), and stimulated −/+ epidermal growth factor (EGF) [25 ng/ml] (30 min). Whole‐cell lysates (WCL) were immunoblotted as indicated. The arrow indicates mTOR auto‐phosphorylated on S2481.
2. TBK1 is required for mTORC1 signaling in MEFs with reduced Akt S473 phosphorylation. TBK1 expression was reduced using lentivirally delivered TBK1 shRNA in Rictor^−/− MEFs stably expressing vector control (V). Cells were EGF stimulated as above, except Torin1 [100 nM] (T) was used to inhibit mTOR.