@inbook{e36e4efa82364dd5aca9f94500b07de9,
title = "Cell-Based Assays Using Primary Endothelial Cells to Study Multiple Steps in Inflammation",
abstract = "Cell-based assays are powerful tools for drug discovery and provide insight into complex signal transduction pathways in higher eukaryotic cells. Information gleaned from assays that monitor a cellular phenotype can be used to elucidate the details of a single pathway and to establish patterns of cross talk between pathways. By selecting the appropriate cell model, cell-based assays can be used to understand the function of a specific cell type in a complex disease process such as inflammation. We have used human umbilical vein endothelial cells to establish three cell-based, phenotypic assays that query different stages of a major signaling pathway activated in inflammation. One assay analyzes the tumor necrosis factor α (TNFα)-induced translocation of the transcription factor NF-κB from the cytoplasm into the nucleus 20 min after stimulation with TNFα. Two more assays monitor the expression of E-selectin and VCAM-1, 4 and 24 h after stimulation with TNFα. Indirect immunofluorescence and high-throughput automated microscopy were used to analyze cells. Imaging was performed with the IN Cell Analyzer 3000. All assays proved to be highly robust. Z′ values between 0.7 and 0.8 make each of the three assays well suited for use in high-throughput screening for drug or probe discovery.",
author = "Thomas Mayer and Bernd Jagla and Wyler, {Michael R.} and Kelly, {Peter D.} and Nathalie Aulner and Matthew Beard and Geoffrey Barger and Udo T{\"o}bben and Smith, {Deborah H.} and Lars Brand{\'e}n and Rothman, {James E.}",
note = "Funding Information: This work was supported by MLSCN Grant 1U54 H6003914‐01 to J.E.R and NIH Grants 1RO3 MH076344‐01, 1RO3 MH076509‐01, and 1RO3 MH076343‐01 to support assay development to T.M. We thank Dr. Ouerfelli (MSKCC) for providing the siRNA. We thank Dr. Martin Wiedmann for his suggestions.",
year = "2006",
doi = "10.1016/S0076-6879(06)14015-X",
language = "English (US)",
isbn = "0121828190",
series = "Methods in Enzymology",
publisher = "Academic Press Inc.",
pages = "266--283",
booktitle = "Measuring Biological Responses with Automated Microscopy",
}