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题目:: Transcription profiling ofrat cortical cells transfected with Math2 reveals Prg1 is regulated by the basic helix-loop-helix transcription factor Math2
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状态:: 发布时间Oct. 21, 2008 , 更新时间 May 1, 2014 , 提交时间 June 19, 2008,
物种:: Rattus norvegicus
摘要:: Math2 (NEX-1/NeuroD6) is a member of the bHLH transcription factor family and is involved in neuronal differentiation and maturation. In the present study, we identified the genes targeted by Math2 using DNA microarrays and cultured rat cortical cells transfected with Math2. Of the genes regulated by Math2, we focused on plasticity-related gene 1 (Prg1). Prg1 expression induced by Math2 was confirmed in cultured rat cortical cells and PC12 cells analyzed by real-time quantitative PCR. Examining the promoter region of rat Prg1, we found four E-boxes designated -E1 to -E4 (CANNTG) which were recognized by the bHLH transcription factor. Using chromatin immunoprecipitation (ChIP) assays, we found that Math2 directly bound to the E-box(es) in the Prg1 promoter. The reporter assay of Prg1 showed that -E1 was critical for the regulation of the Prg1 expression by Math2. Then, the functional role of Math2 and Prg1 was investigated in PC12 cells. Seventy-two hours after transfection of Math2 or Prg1, neurite length and number was significantly induced in PC12 cells. Co-transfection with Prg1-siRNA completely inhibited Math2-mediated morphological changes. Our results suggest that Math2 directly regulates Prg1 expression and Math2-Prg1 cascade plays an important role in neurite outgrowth in PC12 cells. Experiment Overall Design: We identified the genes targeted by Math2 using DNA microarrays and cultured rat cortical cells transfected with Math2.Total RNA was used for cDNA synthesis with a Two-Cycle Target Labeling Kit. Gene expression was analyzed by an Affymetrix GeneChip® Rat Genome 230 2.0 array which was spotted with 31,100 probe sets. Sample preparation for array hybridization was carried out as described in the manufacture’s instructions.
实验种类:: transcription profiling by array
样本量:: 6
实验设计:
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数据号:: E-GEOD-11829, GSE11829
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