Cell culture and transfection

MBE were a gift from R. Auerbach (University of Wisconsin, USA). MBE were cultured in DMEM high glucose (Lonza, Switzerland) supplemented with 10% foetal bovine serum (FBS) (Biowest, France) in a humidified atmosphere at 37°C with 5% CO₂. The cells were routinely checked for mycoplasma contamination by PCR as described previously (Kochan et al., 2016). MBE were seeded in 24-well plates (5×10⁴ cells in 0.5 ml medium per well) and transfected with indicated vectors using Lipofectamine 2000 (Invitrogen) according to the manufacturer's instruction. Where indicated, the cells were stimulated with 10 ng/ml rhIL-1β (PromoKine, Germany), 10 ng/ml rhTNF (PromoKine), 10 ng/ml rmIFNγ (PromoKine), 100 ng/ml PMA (Sigma-Aldrich) or 100 ng/ml LPS (Sigma-Aldrich). Where required, the inhibitors: 5 µM IKK Inhibitor VII (Merck, Germany) or 10 µM U0126 (Cell Signaling Technology) were added 60 min prior to stimulation.