Fig. 3Figure 3
NF-κB p65 interacts with β-catenin and reduces β-catenin-dependent gene transcription in brain endothelium. A-C , Immunofluorescence staining of active β-catenin (non-phosphorylated) in bEnd.3 cells treated with recombinant Wnt3a protein (100 ng/ml) only or Wnt3a protein plus TNF-α (100 ng/ml) or IL-1β (10 ng/ml) for 24 h and its quantifications presented as total signal intensity ( B ) and nuclear signal intensity ( C ). D-E , Immunofluorescence staining of NF-κB p65 in bEnd.3 cells treated with TNF-α or IL-1β and its quantifications presented as nuclear intensity. F , RT-qPCR analysis of the expression levels of Wnt target gene Axin2 in bEnd.3 cells when NF-κB p65 was knocked down and the cells were treated with Wnt3a with or without TNF-α or IL-1β. G , TOP-Flash assay of bEnd.3 cells treated with NF-κB pathway inhibitor BAY 11-7085 (10 mM) and the cells were treated with Wnt3a with or without TNF-α or IL-1β. H , Bimolecular fluorescence complementation (BiFC) assay in HEK 293