Figure 2
DNA methylation analysis of FBN1 CpG island shores of WT and KO alleles in FBN1 het KO fibroblast cells. A diagram of the porcine FBN1 upstream region is illustrated in the upper panel. The SNP between the WT and KO alleles identified in this study is located in the FBN1 CpG island shore. Sequenced sodium bisulfite PCR fragments including the FBN1 CpG island shore were divided into three sub-regions based on methylation status (O, outside the FBN1 CpG island shore. S, shore region. I, CpG island.). The DNA methylation status of each allele in FBN1 het KO fibroblast cells is represented in the lower panel. Depending on the SNP (G or A) present in the sodium bisulfite PCR fragments, sequenced reads were defined as WT (G) or KO (A) alleles for each cell line collected at P6 (early passage) and P20 (late passage). Horizontal and vertical axes indicate the position of the CpG sites (O, 2 CpGs; S, 9 CpGs; I, 13 CpGs) and the number of sequenced reads (29 to 45 reads), respectively. White and black bars indicate unmethylated and methylated CpGs, respectively, consistent with our previous study15. CpGs within the two dotted lines are in the shore region (S) and whose RM allele ratio was calculated in the present study.
