Fig. 5.

Overexpressing RIPK4 activates NF-κB signalling. a NF-κB luciferase reporter activities were analysed in the indicated cells. Each bar represents the mean ± SD of three independent experiments. b EMSA indicating that endogenous NF-κB activity increased in RIPK4-transduced cells and decreased in RIPK4-silenced cells. The OCT-1-DNA binding complex served as a control. c Western blot of nuclear NF-κB-p65, cytoplasmic NF-κB-p65, IKK, p-IKK, IκB and p-IκB levels in the indicated cells. β-actin was used as a loading control. d Immunofluorescence staining showing reduced levels of p65 in shRIPK4 T24 and RT4 cells relative to the control shNC T24 and RT4 cells, and increased levels of p65 in BIU87-RIPK4 cells compared with BIU87-vector cells. *P < 0.05, **P < 0.01 by Student’s t test