Figure 5

Differentiation of the BMP2⁺ cells to cardiac cells. (a) RT-PCR analysis of the cardiac markers in BMP2⁺ cells and other controls. (b) Schematic outline of the protocol used to derive cardiomyocytes from BMP2⁺ cells. (c) The morphology of the contracting EBs. The red arrows indicate the contracile area(s) in that EB. (d) RT-PCR analysis of the representative cardiac markers in 12 day secondary EBs derived by the hanging drop protocol from single cell suspension obtained from seven-day-old primary EBs generated by the protocol outlined in Figure 1 from CGR8 wild-type EBs without puromycin treatment, β-actin puro EGFP EBs and BMP2 EBs with puromycin treatment (videos of the beating clusters in these populations are provided as Additional data files 10-12). (e) Graph showing the relative expression levels of the genes presented in (a) as obtained by Affymetrix analysis. The expression levels of each gene were normalized with its maximum level set as 100%. Each result is an average of three independent experiments (Additional data file 13).