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. 2017 Nov 17;37(1):19–38. doi: 10.15252/embj.201696164

Figure 2. TBK1 promotes growth factor‐induced mTORC1 signaling in a stimulus‐selective manner.

Figure 2

  1. EGF increases mTORC1 signaling in a TBK1‐dependent manner. TBK1+/+ and TBK1−/− MEFs were serum‐starved (20 h), pre‐treated with Ku‐0063794 [1 μM] (30 min), and stimulated −/+ EGF [25 ng/ml] (0–60 min). Whole‐cell lysates (WCL) were immunoblotted (IB) as indicated.
  2. Ectopic expression of TBK1 rescues mTORC1 signaling in TBK1−/− MEFs. TBK1−/− MEFs were transiently transfected with vector control (V), wild type (WT), or kinase‐dead (KD) Flag‐TBK1, serum‐starved (20 h), and analyzed as above.
  3. EGF‐receptor signaling remains intact in TBK1−/− MEFs. TBK1+/+ and TBK1−/− MEFs were serum‐starved (20 h) and stimulated −/+ EGF [25 ng/ml] (0, 5, or 15 min).
  4. TBK1 is required for EGF‐stimulated mTORC1 and mTORC2 signaling. TBK1+/+ and TBK1−/− MEFs were stimulated with EGF as in (A).
  5. TBK1 is not required for insulin‐stimulated mTORC1 signaling. TBK1+/+ and TBK1−/− MEFs were stimulated with insulin as in (C).
  6. Side‐by‐side comparison of EGF‐ vs. insulin‐stimulated mTORC1 and mTORC2 signaling in TBK1+/+ vs. TBK1−/− MEFs. MEFs were treated as in (C).
  7. Pharmacologic TBK1 inhibition reduces EGF‐induced mTORC1 signaling. HEK293 cells were serum‐starved (20 h), pre‐treated with Ku‐0063794 [1 μM] (30 min) or amlexanox [50 μM] (2 h), and stimulated −/+ EGF [25 ng/ml] (30 min) or insulin (INS) [100 nM] (30 min).
  8. EGF but not insulin increases mTOR P‐S2159. TBK1+/+ vs. TBK1−/− MEFs: MEFs were treated as in (C). The arrow indicates mTOR phosphorylated on S2159.

Source data are available online for this figure.