Bms345541

The kinase inhibitor library, composed of 149 selective or broad-spectrum kinase inhibitors dissolved in DMSO (10 mM stock concentration), was purchased from Cayman Chemicals (#10505). The following reagents were used for cell treatments at given concentrations: LPS (Escherichiacoli O55:B5, #L2880, Sigma-Aldrich, 100 ng ml⁻¹), recombinant human TNFα (rHuTNF, #50435.50, Biomol, 10 ng/ml), Birinapant/SM (#HY-16591, MedChem Express, 1 µM), pan-caspase inhibitor zVAD-fmk (#4026865.0005, Bachem, 25 μM), MK2 inhibitor PF3644022 (#4279, Tocris, 5 μM), RIPK1 inhibitor Nec-1 (#BML-AP309-0020, Enzo Life Sciences, 50 µM), RIPK1 inhibitor Nec-1s (# 10-4544-5 mg, Tebu-Bio, 50 µM), RIPK3 inhibitor GSK872 (#HY-101872, MedChem Express, 5 µM), IKKβ inhibitor BMS345541 (#Axon 1731, Axon Medchem, 5 µM), TBK1/IKKε inhibitor BX795 (#T1830, Tebu-Bio), 5-Iodotubercidin (#HY-15424, MedChem Express, 2.5–10 µM), ABT-702 dihydrochloride (#HY-103161, MedChem Express, 5 µM), Staurosporine (#81590, Cayman, 5 µM), Etoposide (#E1383, Sigma, 5 µM), Doxorubicin (#15007, Cayman, 5 µM), Gemcitabine (#S1714, Selleckchem, 100 nM).

All reagents were purchased from Sigma-Aldrich (Zwijndrecht, The Netherlands) unless stated otherwise. In the present study, lipopolysaccharide (LPS) from Escherichia coli 0127:B8 was used. The toxins, p-cresyl glucuronide (pCG) and p-cresyl sulfate (pCS), were synthesized by the Institute for Molecules and Materials, Radboud University, Nijmegen, The Netherlands, as previously described [36 (link)]. IκB kinase (IKK) inhibitor BMS-345541 was obtained from Axon Medchem (Groningen, The Netherlands). Water (LC-MS grade), acetonitrile (ACN; HPLC-S grade) and methanol (HPLC grade) were obtained from Biosolve (Valkenswaard, The Netherlands). Formic acid (analytical grade) was purchased at Merck (Darmstadt, Germany). Ultrapure water was produced by a Milli-Q^® Advantage A10 Water Purification Systems (Merck, Schiphol-Rijk, The Netherlands). Cell culture plates were obtained from Greiner Bio-One (Monroe, NC, USA).