Fig. 4

Rescued 3abKO cells are inefficient at re-establishing imprints on the KvDMR gDMR suggesting a unique regulatory mechanism associated with this imprinted locus. a Methylation level of KvDMR in various ES cell lines by pyrosequencing. The gDMR exhibits significant loss of methylation in 1KO and 3abKO cells, and methylation remains unchanged in both rescued lines. b Validation of pyrosequencing results by COBRA; TaqaI was used to digest the PCR product: me, fragments resulting from cleavage of methylated product; un, fragments when unmethylated. There is no increase in methylated fragments from 3abKO to 3abKO + 3a2 samples. c Clonal analysis for KvDMR in WT, 1KO and 3abKO + 3a2 cells. d Graphical summary of clonal data from c. Error bars indicate s.e.m