Increased canonical NF-κB signaling in patients with nonsense GOF NFKB2 mutation. (A) HEK293T cells growing in 24-well plates were cotransfected with pGL4.32-NF-κB and indicated NFKB2 expression vector. Next day, cells were treated with TNFα for 5 hours and luciferase activities were measured. Western blot analysis confirmed similar protein expression levels. The data were shown as fold change compared with TNFα-treated empty vector (pcDNA3-HA). Results shown are means + standard error of the mean of 4 experiments. *P = .0267, ***P = .0005 (Student t test). (B) Total PBMCs were stimulated with LPS (100 ng/mL) for 24 hours. Cytokine expression in supernatants was analyzed via multiplex bead assay (Luminex). Data shown are from 3 experiments with a heathy donor and indicated patients tested in pairs.