Fig. 5

Decreased Grin2a gene expression correlates with dynamic regulation of Grin2a gene promoter histone modifications. a Relative quantification (2^-∆∆Ct) of Grin2a mRNA levels at 5 different time points (3 h, 7 h, 24 h, 3 days and 2 weeks) after glutamate treatment compared to time-matched sham controls. b Schematic presentation of Grin2a 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 Grin2a at 5 different time points (3; 7; 24 h, 3 days; 2 weeks) after transient glutamate stimulation. Data are expressed as mean fold change over time-matched sham controls after normalization to positive control region of the corresponding antibody. d Bisulfite sequencing of the Grin2a promoter identified increased DNA methylation in glutamate-treated neuronal cultures compared to time-matched sham controls. Positions of analyzed loci relative to the most downstream TSS and p-values of Fisher’s exact test of significant locus specific differences in methylation are show. All error bars represent standard deviation. Asterisks indicate significance (p < 0.05)