Panels A and B: RAW 264.7 cells were treated with γTE at 20 or 40 μM for 16 h (A) or at 20 μM for 8 or 16 h (B). Panel C: BMDMs were treated with γTE at 10 μM for 8 h. Panel D: Subconfluent A549, PC-3 and MCF-7 cells were pre-treated with γTE (20 μM) for 14–16 h, and then stimulated with TNFα for 5 min. Panel E: Sub-confluent A549, PC-3 and MCF-7 cells were pre-treated with γTE (20 μM) with or without subsequent TNFα stimulation (10 ng/ml, 5 min). In these studies, total proteins were immunoblotted with antibodies for CYLD or A20 and β-actin, and cytosolic proteins were analyzed for phospho-IκBα. These data are representative results from at least three independent experiments.