Fig. 5. The Nkx2-5Y-A allele impairs cardiac differentiation in EBs. (A) FACS profiles of dissociated wild-type and Nkx2-5^GFP/+ EBs showing side scatter (SSC-H) versus GFP fluorescence (FL1-H). Gated cells were collected for expression profiling. (B) Comparison of GFP fluorescence versus cell counts in EBs of different genotype. (C) Comparative RT-PCR analysis of gene expression in FACS-purified GFP+ cells from EBs of genotypes indicated. (D) Real-time RT-PCR analysis of gene expression from FACS-purified GFP+ cells as in C. Results reported as percent expression relative to Nkx2-5^GFP/+(N) cells (two independent experiments performed in duplicate; error bars indicate ±s.e.m.).