Fig. 4

Decreased Gria2 gene expression correlates with dynamic regulation of Gria2 gene promoter histone modifications. a Relative quantification (2^-∆∆Ct) of Gria2 mRNA levels at 5 different time points (3 h, 7 h, 24 h, 3 d and 2 weeks) after glutamate treatment compared to control treatment. b Schematic presentation of Gria2 gene promoter region and amplicon localization for qPCR of immunoprecipitated DNA and bisulfite PCR used for DNA methylation analysis. c Chromatin immunoprecipitation of histone modifications H4ac, H3K4me3, H3K27me3 and H3K9me3 at the promoter region of Gria2 at 5 different time points (3 h, 7 h, 24 h, 3 d; 2 weeks) after transient glutamate stimulation. Data are expressed as mean fold change over control treatment plus standard deviation after normalization to positive control region of the corresponding antibody. d Bisulfite sequencing of the Gria2 promoter identified increased DNA methylation of glutamate-treated neuronal cultures compared to sham controls at single CpG (black dots) and non-CpG (red dots) positions. Positions of analyzed loci relative to TSS, non-CpG sequences and p-values of Fisher’s exact test of significant locus specific differences in methylation are shown. All error bars represent standard deviation. Asterisks indicate significance (p < 0.05)