Fig. 3.

Cell proliferation only observed in CHIR99021-treated cardiomyocytes derived from cdh2^−/− mouse ES cells. a Representative FACS plots illustrating purification of VCAM-1-positive mouse cardiomyocytes. b Graphical representation showing fold change in cardiomyocytes proliferation in untreated control and cardiomyocytes treated with N-cadherin antibody (1:1000) or CHIR99021 (1 μM) normalized to control. Error bars indicate s.d., n = 3 replicates. **P < 0.01 for Kruskal-Wallis one-way analysis of variance compared to control. c, d Quantitative PCR analysis of mRNA isolated from cardiomyocytes in untreated control and cardiomyocytes treated with either N-cadherin antibody (1:1000) or CHIR99021 (1 μM). The relative gene expression levels showed that cyclin genes such as Ccnb1, Ccnb2, and Ccnd2 were significantly upregulated only in cardiomyocytes treated with CHIR99021. Correspondingly, genes in canonical Wnt signaling pathway (Wnt1, Wnt2, and Wnt6) and effector genes (Axin2 and Lef1) were shown to be significantly upregulated in cardiomyocytes treated with CHIR99021. Error bars indicate s.d., n = 3 replicates. *P < 0.05 and **P < 0.01 for Kruskal-Wallis one-way analysis of variance compared to control