Bisulfite analysis and Runx1 expression in day 6 EB subpopulations. (A) Methylation patterns of the Runx1 +23 enhancer in c-Kit⁺CD41⁻ and c-Kit⁺CD41⁺ cells from day 6 EBs. Sequencing reactions of individual amplicons are represented by each row of circles. Open circles denote unmethylated CpGs, and filled circles represent methylated CpGs. (B) Methylation patterns of the Runx1 P1 promoter in c-Kit⁺CD41⁻ and c-Kit⁺CD41⁺ cells from day 6 EBs. (C) Quantitative reverse transcriptase polymerase chain reaction analysis of Runx1 P1 and P2 mRNA isoforms in c-Kit⁺CD41⁻ and c-Kit⁺CD41⁺ cells from day 6 EBs. ns, not significant.