使用白细胞的整合素依赖性RhoB激活测定
Integrin Dependent RhoB Activation Assay Using Leukocytes.
作者信息
Lehtonen Janne, Potrzebowska Katarzyna, Samuelsson Malin, Svensson Lena
机构信息
Department of Experimental Medical Science, Lund University, Lund, Sweden.
The School of Medical Sciences, Örebro University, Örebro, Sweden.
出版信息
Bio Protoc. 2018 Dec 5;8(23):e3105. doi: 10.21769/BioProtoc.3105.
To be able to migrate, leukocyte needs to re-use its adhesion molecules to move forward. These adhesion molecules are called integrins and are intracellularly transported via endocytosis and exocytosis in order to translocate to a new site on the cell membrane. The intracellular transportation is regulated by different small GTPases including RhoB. Here we describe an activation assay of RhoB in leukocytes migrating on ICAM-1Fc coated dishes using commercially available Rhoteikin coated agarose beads. Although this is a specific protocol for LFA-1 induced RhoB activation, it can also be used for RhoB activation induced by other soluble and insoluble factors.
为了能够迁移，白细胞需要重新利用其黏附分子来向前移动。这些黏附分子被称为整合素，通过胞吞作用和胞吐作用在细胞内进行运输，以便转移到细胞膜上的新位点。细胞内运输由包括RhoB在内的不同小GTP酶调节。在这里，我们描述了一种使用市售的Rhoteikin包被的琼脂糖珠，在ICAM-1Fc包被的培养皿上迁移的白细胞中RhoB的激活测定方法。虽然这是一种用于LFA-1诱导的RhoB激活的特定方案，但它也可用于其他可溶性和不可溶性因子诱导的RhoB激活。
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