FIGURE 5.

Effects of prenatal caffeine exposure (PCE) on histone acetylation level of 11β‐hydroxysteroid dehydrogenase 2 (11β‐HSD2) and its intrauterine programming mechanism in male offspring. (a) ChIP assay of the histone acetylation level in 11β‐HSD2 promoter region on gestational day 20 (n = 8 per group); (b) ChIP assay of the histone acetylation level in 11β‐HSD2 promoter region at postnatal week (PW) 12 (n = 8 per group); (c) analysis of 11β‐HSD2 promoter region; (d) RT‐qPCR analysis of glucocorticoid receptor (GR) mRNA expression in fetal bone tissue (n = 8 per group); (e) representative immunohistochemical images and quantitative analysis of GR protein in fetal bone tissue (n = 5 per group); (f) the correlation analysis between serum CORT concentration and bone GR protein level on GD20; (g) detection of GR binding to 11β‐HSD2 promoter region through ChIP‐PCR (n = 8 per group); (h) RT‐qPCR analysis of gene expression of HDACs in fetal bone tissue (n = 8 per group). Mean ± SEM, *P < 0.05 compared with the control