Cell experiment:

Cell proliferation experiments are carried out in a 96-well format (6 replicates), Panc 02.13 cells are plated at a density of 2,000 to 5,000 cells per well. At 24 hours following cell seeding, the cells are treated with the tool inhibitor titrations (e.g., TBK1/IKKε-IN-2, 1 nM, 10 nM, 100 nM, 1 μM and 10 μM) for 4 days at 37℃ and then assayed by using the ATP CellTiter-Glo luminescent cell viability assay[1].

TBK1/IKKε-IN-2 is a dual TBK1 and IKKε inhibitor.

TBK1/IKKε-IN-2 (Cmpound #1) inhibits TBK1 biochemical function in Ulight kinase assay at 5 and 250 μM ATP concentration with IC50s of 0.6 and 2.6 nM, rsespectively. TBK1/IKKε-IN-2 inhibits IKKε biochemical function in Ulight kinase assay at 10 μM ATP with an IC50 of 3.9 nM. The IC50 of TBK1/IKKε-IN-2 in the Panc 02.13 proliferation assay is 5 μM[1].

References:
[1]. Muvaffak A, et al. Evaluating TBK1 as a therapeutic target in cancers with activated IRF3. Mol Cancer Res. 2014 Jul;12(7):1055-66.