ABSTRACT
AIM: The main purpose of this study is to investigate the regulatory role of C/EBPα in mouse vesicular glutamate transporter 2(mVGLUT2) gene expression. METHODS: The promoter region of mVGLUT2 was cloned to PGL3-basic vector. Site-direction mutation was used to identify the CCAAT-enhancer-binding protein α(C/EBPα) binding site. The promoter activity was observed by luciferase system. The binding between C/EBPα protein and mVGLTU2 promoter region was determined by EMSA. The C/EBPα gene expression was inhibited by its specific siRNA. RESULTS: mVGLUT2 promoter activity decreased about 50% after mutation of C/EBPα binding site. EMSA showed that C/EBPα protein bound onto mVGLUT2 promoter region. Meanwhile, when C/EBPα gene expression was inhibited by its specific siRNA, mVGLUT2 promoter activity, mRNA level and protein level were decreased about 60%, 40% and 45%, respectively. CONCLUSION: C/EBPα is involved in the regulation of mVGLUT2 gene expression.
ABSTRACT
AIM:The main purpose of this study is to investigate the regulatory role of C/EBP? in mouse vesicular glutamate transporter 2(mVGLUT2) gene expression. METHODS:The promoter region of mVGLUT2 was cloned to PGL3-basic vector. Site-direction mutation was used to identify the CCAAT-enhancer-binding protein ?(C/EBP?) binding site. The promoter activity was observed by luciferase system. The binding between C/EBP? protein and mVGLTU2 promoter region was determined by EMSA. The C/EBP? gene expression was inhibited by its specific siRNA. RESULTS:mVGLUT2 promoter activity decreased about 50% after mutation of C/EBP? binding site. EMSA showed that C/EBP? protein bound onto mVGLUT2 promoter region. Meanwhile,when C/EBP? gene expression was inhibited by its specific siRNA,mVGLUT2 promoter activity,mRNA level and protein level were decreased about 60%,40% and 45%,respectively. CONCLUSION:C/EBP? is involved in the regulation of mVGLUT2 gene expression.