title: [Subcellular localization of human endothelial-overexpressed lipopolysaccharide-associated factor 1 protein].
abstract: OBJECTIVE
  To study the subcellular localization of human endothelial-overexpressed lipopolysaccharide-associated factor 1 (EOLA1) protein in endothelial cells.
  
  
  METHODS
  Human umbilical vein endothelial cell strain ECV304 were cultured in vitro. The fusion protein of enhanced green fluorescent protein (EGFP)-EOLA1 expressing plasmid was constructed. Empty plasmid with EGFP at N side (pEGFP-N2) and fusion protein expressing plasmid EGFP-EOLA1 was respectively transfected into ECV304 cells with liposome. After being cultured for 48 hours, the expression levels of EGFP and fusion protein EGFP-EOLA1 in cells were detected with Western blot. The subcellular localization of EOLA1 protein was detected by laser scanning confocal microscope and immunoelectron microscopy.
  
  
  RESULTS
  The EGFP-EOLA1 coexpression plasmid was verified to be successfully constructed by enzyme cutting and gene sequencing. The fusion protein of EGFP-EOLA1 was observed to express in transfected cells through Western blot. Green fluorescence scattered all over the ECV304 cells transfected with empty plasmid and cells transfected with fusion protein expressing plasmid, and it gathered obviously in the nuclei in the latter cells. Immune deposits were observed in the matrix of cells transfected with fusion protein expressing plasmid but not in the cells transfected with empty plasmid.
  
  
  CONCLUSIONS
  EOLA1 protein is localized in the nucleus and the matrix of ECV304 cell, and it plays its role as a signal transduction factor.
authors.affiliation: Department of Endocrinology, Southwest Hospital, the Third Military Medical University, Chongqing 400038, China.
authors.forename: Min
authors.initials: M
authors.lastname: Luo
authors.forename: Zi-Wen
authors.initials: ZW
authors.lastname: Liang
authors.forename: Zong-Cheng
authors.initials: ZC
authors.lastname: Yang
authors.forename: Xiang-Dong
authors.initials: XD
authors.lastname: Luo
mesh_terms: D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism*; D006801:Humans; D008070:Lipopolysaccharides / Q000378:metabolism; D008565:Membrane Proteins / Q000378:metabolism*; D015398:Signal Transduction
chemical_list: C495142:EOLA1 protein, human; D008070:Lipopolysaccharides; D008565:Membrane Proteins