NF-κB luciferase reporter assays for the IKKγ Δ254–419 and Δ271–419 truncation mutants and D242R mutant.
A, Western blot analysis of IKKγ expression level in IKKγ knock-out human T-cells JM4.5.2, parental Jurkat cells and IKKγ wild type or mutant reconstituted JM4.5.2 cells. B, NF-κB activation was measured using the BrightGlo Luciferase assay system 48 h after transduction with KSHV vFLIP LV (MOI = 50). Bars represent mean fold induction values ± S.D. C, Western blot analysis of IKKγ expression levels in IKKγ knock-out mouse PreB cell line 1.3E2, parental cells 70Z/3, and 1.3E2 cells reconstituted with IKKγ wild type; Δ254–419 mutant or IKKγ D242R. D, NF-κB activation was measured using the BrightGlo Luciferase assay system, 6 h after stimulation with LPS (10 μg ml−1) or 48 h after transduction with KSHV vFLIP LV. Bars represent mean of relative luminescence unit (RLU) values ± S.D.