Figure 2

Activation of NF-κB through NRAGE is specific to the non-canonical BMP pathway. A: 293 cells were transfected with full length NRAGE:EGFP or stimulated with BMP-4 and incubated with or without IKK VII inhibitor, and NF-κB activation assessed by luciferase assay. B: 293 cells were transfected with full length NRAGE:EGFP or stimulated with BMP-4 and incubated with or without Tak1 or Tab1 morpholinos, and NF-κB activation assessed by luciferase assay. C: Endogenous phosphorylation of IKK-α/β in 293 cells transfected with Tak1 or Tab1 morpholinos prior to 10 ng/ml BMP-4 stimulation for one hour. D: 293 cells were transfected with full length NRAGE:EGFP, incubated with or without Tak1 or Tab1 morpholinos immunoprecipitated with rabbit anti-GFP and western blotted for IKK α/β-phosphorylation and total IKK α. E: Western blot of Tak1 and Tab1 proteins in 293 cells with and without morpholinos. Luciferase assays were performed in triplicate and presented as fold increase over renilla. * P < 0.05; vs NRAGE construct or BMP-4 treatment by the two-tailed unpaired Student's t-test.