Effect of ischemia or hypothermia on methylation regulating factors (A). We measured the mRNA levels of methylation related proteins such as DNA methyltransferases (DNMTs) 1, 2, 3, and 4 using real-time PCR. RNA was harvested from cells exposed to 4 hours of oxygen glucose deprivation (OGD) and to reperfusion for 3 to 24 hours. (B) The levels of methyl-CpG binding domain proteins (MBDs) 1, 2, 3, and 4 along with methyl-CpG binding protein-2 (MECP-2) were measured using the same samples and analysis tools as in 6A. Values represent the mean ± S.E.M. of three different experiments.