Technical Reference #1663

1663.

ERM protein moesin is phosphorylated by advanced glycation end products and modulates endothelial permeability Xiaohua Guo; Lingjun Wang; Bo Chen; Qiang Li; Jiping Wang; Ming Zhao; Wei Wu; Ping Zhu; Xuliang Huang; and Qiaobing Huang, Southern Medical Universirt, American Journal of Physiology: Heart Circulatory Physiology, 297(1663), (2009)
Link To Paper

Abstract:
Advanced glycation end products (AGEs) accumulated in different pathological conditions have the potent capacity to alter cellularproperties that include endothelial structural and functional regulations.

Keywords:
vascular permeability; receptor for advanced glycation end products; Rho kinase; mitogen-activated protein kinase; ezrin/radixin/moesin

Materials & Methods:
Fluorescent staining. ECs were plated in gelatin-coated glassbottom microwells (MatTek) and cultured to confluence. After the appropriate treatments cells were fixed and permeated for 15 min at room temperature in PBS with 3.7% formaldehyde and 0.5% Triton X-100. Cells were blocked in 5% BSA in PBS for 1 h and subsequently incubated with moesin or p-moesin antibody at room temperature for 2 h. After a thorough wash in PBS cells were stained with FITC-conjugated secondary antibody (Zymed San Francisco CA). For F-actin staining cells were incubated with rhodamine-phalloidin (2000 U/l) for 40 min at room temperature. Finally cells were washed three times with PBS mounted to allow observation and imaged with a Leica TCS SP2 laser confocal scanning microscope (Wetzlar Germany).

Microscopic Technique
Confocal Microscopy, Laser Scanning

Cell Type(s)
HMVECs