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. Author manuscript; available in PMC: 2014 Oct 1.
Published in final edited form as: Cancer Discov. 2014 Jan 20;4(4):452–465. doi: 10.1158/2159-8290.CD-13-0646

Figure 2. CYT387 inhibits JAK and TBK1/IKKε signaling.

Figure 2

A, In vitro kinase assay with His-TBK1 (4 nM) or GST-IKKε (2 nM), 100 µM IKKε-tide, 100 µM ATP, and CYT387. Assays performed in duplicate, ADP generated normalized to DMSO control, mean ± SD shown.

B, Immunoblot of S418 pCYLD, CYLD, TBK1, IKKε, and β-Actin levels in 293T cells 24 h following transient co-transfection of Myc-tagged CYLD with TBK1-WT, TBK1-KD, IKKε-WT, or IKKε-KD and 4 h treatment with DMSO or 5 µM MRT67307, CYT387, or Ruxolitinib.

C, Immunoblot of S396 pIRF3, IRF3, S172 pTBK1, TBK1, IKKε, and β-Actin levels in macrophages stimulated with LPS (100 ng/ml) for 2 h ± DMSO or inhibitor pretreatment at the indicated concentrations for 1 h.

D, mRNA levels of IFNB1, CCL5, and IL6 in macrophages stimulated with LPS (100 ng/ml) for 2 h ± pretreatment with inhibitors for 1 h. Mean and SEM of triplicate samples shown.