Analysis of focal adhesions and cytoskeleton by custom microarray.
Analysis of focal adhesions and cytoskeleton by custom microarray.
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DOI：
10.1007/978-1-59745-353-0_10
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发表时间：
2007
影响因子：
--
通讯作者：
M. Dalby;S. Yarwood
中科院分区：
文献类型：
--
作者：
M. Dalby;S. Yarwood
粘着斑和细胞骨架（中间丝、微丝、微管）参与了机械力转导--既有直接的（将机械力转导到细胞核），也有间接的（将化学信号级联转导到细胞核）。因此，观察粘着斑和细胞骨架组织的变化在药物治疗和体外医疗材料测试等研究中是非常宝贵的。在这里，我们描述了如何对人类成纤维细胞进行粘着斑蛋白（位于粘着斑）、肌动蛋白（微丝）、微管蛋白（微管）和波形蛋白（中间丝）染色，以及如何进行定制的微阵列实验。免疫荧光和阵列数据的比较分析应使研究人员建立一个全球性的图片的变化，直接和间接mechanotransduction通过细胞骨架从局灶性粘连。
Focal adhesions and the cell cytoskeleton (intermediate filaments, microfilaments, microtubules) are involved in mechanotransduction—both direct (transduction of mechanical forces to the nucleus) and indirect (transduction of chemical signaling cascades to the nucleus). Thus, observation of changes in focal adhesion and cytoskeletal organization can be invaluable in research such as drug treatments and medical material testing in vitro.Here we describe how to stain human fibroblasts for vinculin (located to focal adhesions), actin (microfilaments), tubulin (microtubules), and vimentin (intermediate filaments) and how to perform custom microarray experiments. Comparative analysis of the immunofluorescence and array data should allow the researcher to build up a global picture of changes to both direct and indirect mechanotransduction through the cytoskeleton from focal adhesions.