Skip to main content
. 2016 Apr 29;6:24905. doi: 10.1038/srep24905

Figure 5. An active GRE site was responsible for higher CRH promoter activity in tree shrews.

Figure 5

(A,B) TF binding site prediction analysis revealed an active GRE (aGRE) site was located in the CRH promoter region of tree shrew and it was confirmed by ChIP assay using GR antibody and PCR of tree shrew aGRE. (C) Promoter activity analysis revealed that the difference of CRH promoter activity between human and tree shrew diminished along with the depletion of the aGRE site (*P < 0.05, **P < 0.01, n = 3, Two Way Anova). (D) Statistical table showed the CRH promoter activity of mutant tree shrew retained a lower activity than wild type after treatment with dexamethasone. (E) The aGRE mutation caused a significant decrease of luciferase activity compared to wild type under the treatment of dexamethasone or control solvent. Mifepristone + dexamethasone treatment significantly antagonized the inhibition effect of dexamethasone on CRH promoter activities in N2a cells transfected with wild type or aGRE mutation reporter plasmids. (*P < 0.05, **P < 0.01, ###P < 0.001, n = 4, Two Way Anova). (F–I) CRH mRNA expression was increased after the aGRE site (underlined) was introduced into the CRH gene locus by a pair of guide RNAs (blue) using Cas9/CRISPR tech (*P < 0.05, **P < 0.01, n = 3, One Way Anova).