Immunodetection of Human Double Homeobox 4
Immunodetection of Human Double Homeobox 4
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DOI：
10.1089/hyb.2010.0094
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发表时间：
2011-04-01
期刊：
影响因子：
--
通讯作者：
Tapscott, Stephen J.
中科院分区：
文献类型：
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作者：
Geng, Linda N.;Tyler, Ashlee E.;Tapscott, Stephen J.
DUX 4是面肩肱肌营养不良症（FSHD）的候选致病基因，FSHD是最常见的以进行性骨骼肌变性为特征的肌营养不良症之一。尽管在理解FSHD的精确遗传学方面取得了很大进展，但该疾病的分子病理生理学仍不清楚。主要的限制之一是研究DUX 4蛋白的适当分子工具的可用性。在本研究中，我们报告了五个新的单克隆抗体的发展，针对人DUX 4的N-和C-末端，并使用Western印迹和免疫荧光染色表征其反应性。此外，我们表明，典型的完整编码DUX 4的表达诱导人原代肌细胞中的细胞死亡，而DUX 4的较短剪接形式的表达不会导致这种毒性。这些新抗体的免疫染色揭示了两种DUX 4亚型对人类肌肉细胞的差异效应。这些抗体将为研究DUX 4在FSHD发病机制中的作用提供极好的工具。
Double homeobox 4 (DUX4) is a candidate disease gene for facioscapulohumeral dystrophy (FSHD), one of the most common muscular dystrophies characterized by progressive skeletal muscle degeneration. Despite great strides in understanding precise genetics of FSHD, the molecular pathophysiology of the disease remains unclear. One of the major limitations has been the availability of appropriate molecular tools to study DUX4 protein. In the present study, we report the development of five new monoclonal antibodies targeted against the N- and C-termini of human DUX4, and characterize their reactivity using Western blot and immunofluorescence staining. Additionally, we show that expression of the canonical full coding DUX4 induces cell death in human primary muscle cells, whereas the expression of a shorter splice form of DUX4 results in no such toxicity. Immunostaining with these new antibodies reveals a differential effect of two DUX4 isoforms on human muscle cells. These antibodies will provide an excellent tool for investigating the role of DUX4 in FSHD pathogenesis.