Lehtonen Janne, Potrzebowska Katarzyna, Samuelsson Malin, Svensson Lena
Department of Experimental Medical Science, Lund University, Lund, Sweden.
The School of Medical Sciences, Örebro University, Örebro, Sweden.
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。
