Expression data of MED12 full-length and domain deleted construct overexpression and RNAi knockdown in HEK293 cells.

MED12 is an X-chromosome member of the Mediator complex that is a key regulator of tissue specific gene expression and moderates intracellular signaling via multiple developmental pathways. Sequence variations in the carboxy-terminus of MED12, which contains a PQL and Opa domain, are associated with X-linked mental retardation behavioral syndromes and schizophrenia. Unfortunately, the mechanism(s) through which MED12 sequence variation in the carboxy-terminus could alter vulnerability to neurodevelomental and neuropsychiatric illnesses is yet unclear. In order to elucidate a better understanding of this process, we examined the role of the MED12 carboxy-terminus in cell cycle and gene expression with full-length and domain deleted overexpression constructs and RNA interference in HEK293 cells. Our microarray data show a set of genes differentially expressed in the experimental conditions versus the GFP control. The top 50 most differentially expressed genes in the experimental conditions versus the GFP control also show that MED12 expression level differentially affects stress response and transcriptional regulation pathways. These results are consistent with prior studies showing that MED12 has a key role in determining neuronal cell fate and our theoretical understanding of the biological basis of psychosis. They also lend further insight upon the pathways through which MED12 exerts its effects upon differentiation and disease pathogenesis, which may one day lead to new approaches to the treatment of MED12-related disorders. 12 samples were analyzed, being comprised of four conditions with three biological replicates. Comparisons were made between the GFP control to experimental condition (i.e. GFP vs MED12 FL; GFP vs MED12 PQL/Opa; and GFP vs MED12 shRNA 5)

MED12是中介体复合物（Mediator complex）的X染色体编码成员，该复合物是组织特异性基因表达的关键调控因子，可通过多条发育通路调控细胞内信号转导。MED12的羧基端含有PQL结构域与Opa结构域，该区域的序列变异与X连锁智力障碍行为综合征及精神分裂症密切相关。遗憾的是，目前尚不清楚MED12羧基端的序列变异通过何种机制改变个体对神经发育和神经精神疾病的易感性。为深入阐明这一过程的机制，我们借助全长及结构域缺失的过表达构建体，联合RNA干扰（RNA interference）技术，在HEK293细胞中探究了MED12羧基端在细胞周期调控与基因表达中的作用。本研究的微阵列（microarray）数据显示，与绿色荧光蛋白（GFP）对照组相比，实验条件下存在一批差异表达基因。实验条件与GFP对照组间排名前50的差异表达基因同样表明，MED12的表达水平可差异性影响应激反应与转录调控通路。上述结果与既往研究相符：既往研究证实MED12在决定神经元细胞命运中发挥关键作用，同时也契合我们对精神疾病生物学基础的理论认知。本研究进一步揭示了MED12影响细胞分化与疾病发病机制的具体通路，未来或可为MED12相关疾病的治疗提供全新策略。本次研究共分析12份样本，涵盖4种实验条件，每种条件设置3次生物学重复。实验设置了GFP对照组与各实验组的比较：即GFP vs MED12全长（FL）；GFP vs MED12 PQL/Opa结构域缺失突变体；以及GFP vs MED12短发夹RNA（shRNA）干扰组5。