Figure 4

Methylation patterns in DMRs of imprinted genes in sperm from offspring of diabetic (OD) and non-diabetic (ON) females. OD (n = 9)and ON (n = 9) (7–8 weeks old) mice were killed and spermatozoa were collected and treated with bisulfite and then amplified by nest-PCR. (A ~ C) The products of PCR were digested by enzymes and analyzed by 2.5% agarose gel electrophoresis. The red arrow heads point to the samples that were only partially digested by enzymes. (D, E) The methylation rates in DMRs of H19 and Gtl2 were evaluated by sequencing. Black circle, methylated; white circle, unmethylated; blank loci, CpG lost.