(A) WT, IKKα KO or IKKβ KO SW620 cells were seeded for 48 h prior to treatment with 10 µM BIX02514 or DMSO vehicle for 30 min before treating with TNFα (10 min) and processing for western blot. (B) WT, IKKα KO or IKKβ KO SW620 cells were transiently transfected with reporter plasmids DNA. The following day, cells were treated with BIX02514 or DMSO vehicle control for 30 min, prior to treatment with TNFα for 2 or 4 h. Firefly luciferase luminescence was normalised as above and expressed as log₂(fold change in TNFα-induced luciferase activity relative to the relevant, matched untreated condition). Results are mean ± SD of three independent experiments, each seeded in technical triplicate. Significance testing was performed using two-way ANOVA (repeated measures) with Tukey post-hoc test. P values relate to comparison with corresponding timepoint(s) of WT control.