Figure 6.

HDAC2-dependent downregulation of histone methyltransferase Ehmt2 (G9a) mediates the loss of repressive histone methylation at the Nedd4-1 promoter in stressed animals. A, B, Immunoblots and quantification analysis of the protein level of Nedd4-1 in PFC punches from control versus stressed rats with the intraperitoneal injection of MGCD0103 versus vehicle control (A) or the local injection of GFP versus HDAC2 shRNA lentivirus (B). **p < 0.01, *p < 0.05, ANOVA. C, Quantitative real-time RT-PCR data on the mRNA level of Nedd4-1 in PFC from control versus repeatedly stressed rats. **p < 0.01, t test. D, Representative blots and quantification showing the ubiquitination of GluR1 subunits in control (con) versus repeatedly stressed (RS) rats with the intraperitoneal injection of vehicle, MGCD0103, or MS-275. Lysates of PFC slices were immunoprecipitated with anti-GluR1 and then blotted with an ubiquitin antibody. **p < 0.01, ANOVA. E, ChIP assay data showing the H3K9Me2 level at rat Nedd4-1 promoter regions in PFC lysates from control versus repeatedly stressed (RS) rats. *p < 0.05, t test. Top, Diagram showing the location of primers. F, Quantitative real-time RT-PCR data on the mRNA level of Ehmt1 and Ehmt2 in PFC from control versus repeatedly stressed rats without or with the intraperitoneal injection of MGCD0103. *p < 0.05, ANOVA. G, Quantitative real-time RT-PCR data on the mRNA level of Ehmt2 in PFC from control versus repeatedly stressed rats with the local injection of GFP or HDAC2 shRNA lentivirus. **p < 0.01, ANOVA. H, ChIP assay data showing the acetylated histone H3 level at rat Ehmt2 promoter regions in PFC lysates from control versus repeatedly stressed (RS) rats. *p < 0.05, t test. Top, Diagram showing the location of primers.