Figure 3

Validation of GR binding sites and effects on mRNA expression. A ChIP-PCR validation of identified GBS, previously shown to be GR-targets in literature (grey bars) or representing newly identified GBS (black bars). The genes that are associated with the GBS are listed on the x-axis. The y-axis represents the % of input DNA that was bound by the GR after subtracting the aspecific IgG-bound fraction and the amount of GR bound after vehicle (VEH) treatment. The error bars represent the standard error of the mean (SEM) when comparing the DEX-induced GBS versus the VEH-induced GBS. An unpaired two-tailed T-test was used for statistics. B Diagram indicating whether the known GR-binding regions were previously detected in other published GR-ChIPseq studies based on BlastZ-based interspecies conservation (http://main.g2.bx.psu.edu/) [37]. The genomic locations corresponding to the GBS are listed in Additional file 3: Table S3 as region numbers 1 (Ddc), 7 (FRMD8), 8 (Per1), 11 (Snx7), 14 (Il20ra), 17 (Th), 75 (TLE3), 94 (Ddit4), 345 (Olr1735), 352 (Fndc7), 366 (Pik3r5), 526 (Cry2), 704 (Nfasc), 842 (Narg2), 976 (Kcnab1), 1020 (Ctsd). C mRNA expression of the genes associated with the validated GBS after DEX-treatment relative to VEH-treatment (100%). Expression was normalized against tubulin 2a mRNA expression. The non-parametric Wilcoxon Signed Ranks Test was used for statistics.