Neonatal promoter hypermethylation associated with gene transcription repression. After HiSeq-BSP, genes with consistent DMRs with RRBS results were subjected to real-time qPCR analysis to determine the differences of their expression levels with respect to prenatal age (PN), neonatal response (NN) and NEC disease (NEC). Four NN-genes (CYP2W1, GPR146, TOP1MT, CEND1), related to intestinal metabolism, were significantly hypermethylated in their promoter CGIs (left), and transcriptionally downregulated in the 4d-preterm compared with 0d-preterm (right). The quantitative ratios are normalized to the expression of GAPDH. The data are representative of three similar experiments and are displayed as the mean ± SD. *, P < 0.01 as evaluated using the Student’s t-test and with a fold change > =2.