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

Figure EV4. TBK1 promotes mTORC1 signaling in RAW264.7 macrophages upon TLR3 and TLR4 activation (related to Figs 5 and 6).

Figure EV4

  1. siRNA‐mediated knockdown of TBK1/IKKε reduces TLR3/4‐stimulated mTORC1 signaling. RAW264.7 macrophages were transfected with siRNAs targeting TBK1 and IKKε. After 5 days, cells were pre‐treated with rapamycin [20 ng/ml] (30 min) and stimulated −/+ poly(I:C) [30 μg/ml] or LPS [100 ng/ml] (60 min). ns = non‐specific band.
  2. Pharmacologic inhibition of TBK1 reduces mTOR catalytic activity as measured by mTOR S2481 auto‐phosphorylation. RAW264.7 macrophages were pre‐treated with the TBK1 inhibitors BX‐795 [10 μM] (30 min), MRT‐67307 [10 μM] (30 min), amlexanox [100 μM] (2 h) or with the mTOR inhibitor Ku‐0063794 [1 μM] (30 min) and stimulated −/+ poly(I:C) [30 μg/ml] or LPS [100 ng/ml] (60 min). WCL was immunoblotted as indicated.