DNA methylation analysis of tumour suppressor genes by bisulfite sequencing. Bisulfite sequencing of RARB, CST6, CCND2 (MCF-7 cells), and CDKN2A (HCC1954 cells) gene promoters after 6 hours reprogramming. Schematics indicate the position of analysed CpG islands in promoter regions. A minimum of 10 clones were analysed for each gene and average loss of methylation was calculated for each reprogramming treatment. Black circles indicate metylated CGs, white circles indicate unmethylated CGs. Reprogramming in AOE produced the highest levels of demethylation (P < 0.05; a = AOE, XOE, ESCE vs UN; b = AOE vs XOE; c = AOE vs ESCE).