Mouse Homologue of the Schizophrenia Susceptibility Gene ZNF804A as a Target of Hoxc8
Figure 2
The mouse Zfp804a gene is located in chromosome 2 and has Hoxc8 responsive elements in the intron1 region. (a) Zfp804a gene organization. Coding regions or exons (black box) and the introns (grey box) are shown. Hoxc8-binding core sequences are shown in bold and the underlined sequences indicate primer targets for ChIP-PCR. (b) A ChIP experiment was performed using mouse E11.5 embryos and F9 cell line. The DNA-Hoxc8 complexes were immunoprecipitated by a Hoxc8-monoclonal antibody (lane 4). Input (prior to immunoprecipitation) was used for an internal control (lane 1), and mouse IgG and vehicle were used as a negative control (lane 3-4). (c) Hoxc8 activates Zfp804a in the effector-reporter assay. Luciferase construct is driven by a minimal promoter fused to the intronic segment of Zfp804a (pGL3-MP : Zfp804a; reporter) and cotransfected with a empty vector pcDNA3, pcDNA3:Hoxc8 (effector), and a siRNA for Hoxc8 in F9 cells. Thirty-six hours after transfection, luciferase activities were measured. The experiments were performed in 3 independent trials. The mean values of the luciferase activity are shown as fold change relative over that of the pcDNA3.
