Figure 1. Inducible activation of Wnt/β-catenin signaling in arterial ECs.

(a) Flow cytometric analysis of ECs. ECs were collected from the heart of Bmx-CreER^T2 mice (Ctrl) or from Bmx-CreER^T2 crossed with CAG-CAT-EGFP mice (Bmx/EGFP) 1 week after the TAM treatment. (b) Immunofluorescent staining of cardiac tissue for GFP (red), CD31 (green), and TO-PRO-3 (blue). Scale bars: 20 μm. (c, d, e) Genotyping PCR (c), western blot (d) and quantitative RT-PCR analysis (e) of cardiac ECs isolated from Ctrl (Bmx-CreER^T2 with Ctnnb1^+/+) mice (Ctrl ECs) and Bmx/CA mice (Bmx/CA ECs). (c) Floxed allelle of β-catenin (=500 bp) was detected in Bmx/CA ECs but not in Ctrl ECs. (d) β-catenin protein lacking exon3 (=75 kDa) was detected in Bmx/CA ECs but not in Ctrl ECs. (e) Expression levels of Wnt/β-catenin signaling target genes (Axin2 and Lef1) were higher in Bmx/CA ECs compared with Ctrl ECs. **P < 0.01.