Figure 2. disiRNA loci have an on/off pattern of DNA methylation.

(A) Bisulfite sequencing result of the ζ-η locus reveals significant methylation in all DNA clones examined. Each row of circles represents the number of cytosines in one subcloned ζ-η fragment. Opened and filled circles indicate unmethylated and methylated cytosines, respectively. (B) Strategy for detecting methylation in disi-6 locus (strategy 1, Materials and Methods). The lack of cleavage by DpnII indicates no methylation at the GATC recognition site of the original sequence, whereas the cleavage by DpnII indicates that the DNA sequence contained 5mC. (C) 9 clones from “cut” and “uncut” populations were subject to sequencing. DNA methylation profiles were shown. (D) Southern blot analyses of disi-29 and disi-47 loci. The am and ζ-η loci were as the negative and positive control, respectively.