Figure 7.

Chromatin immunoprecipitation of CBP, histone H3–K9 acetylation and NGFI-A binding to the exon 1₇ GR and DNA methylation analysis of NGFI-A-bound exon 1₇ GR sequence in hippocampal tissue of 6-d-old high and low LG offspring (n = 4 animals per group). a–c, Lanes were loaded with non-immunoprecipitated input (I), CBP, acetylated histone H3–K9 or NGFI-A primary antibody immunoprecipitated (A), or non-immune IgG antibody immunoprecipitated (N) hippocampal extracts. a, Representative Southern blots of the amplified exon 1₇ region of the GR from NGFI-A immunoprecipitated (IP) hippocampal tissue (194 bp band). Exon 1b estrogen receptor-α promoter region, which does not contain NGFI-A recognition elements (493 bp), amplified from the same NGFI-A immunoprecipitated hippocampal tissue was run as a control for specificity and showed no signal. b, Representative Southern blots of the amplified exon 1₇ region of the GR from CBP immunoprecipitated hippocampal tissue (194 bp band). c, Representative Southern blots of the amplified exon 1₇ region from acetyl-histone H3–K9 immunoprecipitated hippocampal tissue (194 bp band) and β-actin (171 bp band) control. d, ROD (mean ± SEM) of exon 1₇ sequence amplified from cAMP, acetyl-histone H3–K9, or NGFI-A immunoprecipitated hippocampal tissue of 6-d-old high and low LG offspring (n = 4 animals per group; *p < 0.05; **p < 0.001; ***p < 0.0001). e, Mean ± SEM percentage methylation per cytosine for the 5′ and 3′ CpG dinucleotides within the NGFI-A binding region of the exon 1₇ GR promoter bound to NGFI-A protein in 6-d-old offspring of high and low LG mothers (6–10 clones sequenced per animal; n = 4 animals per group; **p < 0.001; ***p < 0.0001).