Fig. 4

Sanger bisulfite sequencing of PCDHA4 promoter region revealed different methylation patterns in PAX3/FOXO1 ARMS cell lines and ERMS cells. Sequencing was performed for at least 8 clones obtained by subcloning bisulfite-converted promoter region. Sequenced region spanned from position +94 to +828, where position +1 corresponds to the gene transcription start site (TSS). The sequence investigated maps on a predicted CpG island and includes the region identified with iChip algorithm. ARMS cell lines have high methylation levels (RH4, RH30) with respect to ERMS cell lines (RH36, RD). Circles: cytosine within CpG dinucleotides; black circles: methylated cytosine; white circles: unmethylated cytosine