Fig. 2Figure 2
LPS and proinflammatory cytokines suppress brain endothelial Wnt/β-catenin signaling. A-B , Co-immunofluorescence staining of active β-catenin (non-phosphorylated) and CD31 in the brain cortex tissue of mice treated with LPS or saline ( A ) and its quantifications ( B ). Active β-catenin signal in endothelial cells was selectively quantified. n = 6 mice per group. ( C-D ) Co-immunofluorescence staining of LEF1 and CD31 in the brain cortex tissue of mice treated with LPS or saline ( C ) and its quantifications ( D ). n = 6 mice per group. E , RT-qPCR analysis of the expression levels of Wnt/β-catenin signaling target genes Axin2 , Apcdd1 , Nkd1 , and Notum in brain endothelial cells (ECs) isolated by flow cytometry from saline or LPS-treated mice (normalized to β-actin). n = 7 mice per group. F-H , RT-qPCR analysis of the expression levels of Axin2 in mouse brain microvascular ECs (bEnd.3) treated with recombinant Wnt3a protein (100 ng/ml) or Wnt3a protein plus LPS ( F )