Journal: BMC Genomics
Article Title: Changes in gene methylation patterns in neonatal murine hearts: Implications for the regenerative potential
Figure Lengend Snippet: qPCR validation of microarray results for remarkable genes showing significant changes in DNA methylation and transcript levels between d1 and d7 in neonatal murine hearts. The left panels show DNA methylation and gene expression microarray results represented by red bars and black markers, respectively; DNA methylation is expressed as MeDIP enrichment and gene expression levels as normalized microarray signals. The black markers preceding that indicating the neonatal d1 represent gene expression levels in embryonic hearts E16, E18, E19, E20. The middle panels present CpG methylation estimated with Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) where the DNA methylation levels correspond to the amounts of DNA undigested by CpG methylation dependent McrBC enzyme (1-(McrBC/Input). The right panels demonstrate transcript levels determined with qPCR as the ratios to the reference transcript of the Tbp gene (TATA binding protein gene). The microarray results were determined for pooled samples of 3 mice. The qPCR results represent average values obtained for three individuals for each developmental time-point. The statistical significance has been determined with two-tailed heteroscedastic Student’s t -test. The complete results of qPCR validation are collected in Additional file 2 : F2
Article Snippet: Validation of CpG methylation CpG methylation levels were examined by using Methylation Dependent Restriction Digestion followed by quantitative PCR (MDRE-qPCR) with McrBC restriction endonuclease (NEB, cat. no. M0272).
Techniques: Real-time Polymerase Chain Reaction, Microarray, DNA Methylation Assay, Expressing, Methylated DNA Immunoprecipitation, CpG Methylation Assay, Methylation, Binding Assay, Mouse Assay, Two Tailed Test