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. 2010 Sep 1;35(13):2521–2537. doi: 10.1038/npp.2010.117

Figure 2.

Figure 2

Several memory/plasticity-related genes present an acetylated-H2B/H4 enrichment on their promoters during the consolidation of a spatial memory. (a) mRNA expression levels of several CBP/memory-related genes: bdnf-eIV, cFos, FosB, and zif268 were evaluated by RT-qPCR in the dorsal hippocampus of control (visible platform, VPf) and learning (hidden platform, HPf) groups trained for 3 days. RNA polymerase II was tested as an internal negative control. Values were normalized to the 18S ribosomal subunit. Data are expressed as mean (±SEM) (n=6 per group), with percentage relative to the control group (VPf) arbitrarily set at 100%. Fold inductions are indicated above histograms. Student's t-test: *p<0.05, as compared with VPf. (b) Chromatin immunoprecipitation were performed on 3-day-trained rats from the VPf and HPf groups (n=3). Semiquantitative PCR were performed to analyze the presence of bdnf-pIV, cFos, FosB, and zif268 proximal promoters in the immunoprecipitates. Controls for immunoprecipitation were performed with an unrelated control antibody (Ct-IgG) or no antibody (noAb). Input DNA corresponding to the total of chromatin before immunoprecipitation was run in parallel as positive control. Representative results are shown in duplicate for each gene tested. β-Actin was used as internal control. Bands were quantified (ImageJ) and fold inductions (f.i.) are indicated in the table below. An activation score was arbitrarily affected by the HPf vs VPf condition: (+++) when f.i. >2.00; (++) when f.i. >1.50; (+) when f.i. >1.25; and (=) when −0.25<f.i.>+0.25. acH3, acetylated K9, K14 H3 histone; acH4, acetylated K12 H4 histone; acH2B, acetylated K12, K15 H2B histone; Bdnf-eIV, bdnf exon IV; bdnf-pIV, bdnf promoter IV.