Jove
Visualize
Contact Us
JoVE
x logofacebook logolinkedin logoyoutube logo
ABOUT JoVE
OverviewLeadershipBlogJoVE Help Center
AUTHORS
Publishing ProcessEditorial BoardScope & PoliciesPeer ReviewFAQSubmit
LIBRARIANS
TestimonialsSubscriptionsAccessResourcesLibrary Advisory BoardFAQ
RESEARCH
JoVE JournalMethods CollectionsJoVE Encyclopedia of ExperimentsArchive
EDUCATION
JoVE CoreJoVE BusinessJoVE Science EducationJoVE Lab ManualFaculty Resource CenterFaculty Site
Terms & Conditions of Use
Privacy Policy
Policies

Related Experiment Videos

Cell-based assays using primary endothelial cells to study multiple steps in inflammation.

Thomas Mayer1, Bernd Jagla, Michael R Wyler

  • 1The Judith P. Sulzberger, MD, Columbia Genome Center, Department of Physiology and Cellular Biophysics, Columbia University, New York, NY, USA.

Methods in Enzymology
|November 18, 2006
PubMed
Summary
This summary is machine-generated.

Related Concept Videos

You might also read

Related Articles

Articles linked to this work by shared authors, journal, and citation graph.

Sort by
Same author

Turbocharging synaptic transmission: 12 SNAREpins are required for rapid release of reconstituted synaptic vesicles.

Science advances·2026
Same author

Wafer-Scale Electrical Characterization of Al/Al<sub><i>x</i></sub>O<sub><i>y</i></sub>/Al Tunnel Junctions for Process Monitoring at Room Temperature.

Nanomaterials (Basel, Switzerland)·2026
Same author

Operating CRISPR/Cas12a in a complex nucleic acid sequence background.

Nucleic acids research·2026
Same author

Stress and conflict management in core facilities-News from an imaging survey.

Journal of microscopy·2026
Same author

Retrospective observational study of treatment and referral pathways of patients with epilepsy in Germany.

Seizure·2026
Same author

'There's No Improvement. I'm Still Bedridden': Recovery After Hospital Admission for Low Back Pain and Alignment With Patient Expectations.

Musculoskeletal care·2026

This study developed three robust cell-based assays using human umbilical vein endothelial cells to analyze inflammation signaling pathways. These assays are suitable for high-throughput screening in drug discovery.

Area of Science:

  • Cell biology
  • Molecular biology
  • Pharmacology

Background:

  • Cell-based assays are crucial for drug discovery and understanding cellular signaling pathways.
  • Phenotypic assays provide insights into complex biological processes like inflammation.
  • Human umbilical vein endothelial cells (HUVECs) are relevant models for studying inflammation.

Purpose of the Study:

  • To establish and validate robust cell-based phenotypic assays for inflammation signaling.
  • To investigate key events in the tumor necrosis factor alpha (TNFα) signaling pathway.
  • To assess the suitability of these assays for high-throughput screening (HTS) in drug discovery.

Main Methods:

  • Development of three distinct cell-based assays using HUVECs.
  • Stimulation with TNFα to activate inflammatory signaling.

Related Experiment Videos

  • Monitoring of NF-kappaB nuclear translocation, E-selectin, and VCAM-1 expression.
  • Utilizing indirect immunofluorescence and automated microscopy (IN Cell Analyzer 3000) for analysis.

    • Main Results:

    • Successfully established three robust assays querying different stages of TNFα-induced inflammation signaling.
    • NF-kappaB translocation observed within 20 minutes post-TNFα stimulation.
    • E-selectin and VCAM-1 expression monitored at 4 and 24 hours post-TNFα stimulation.
    • High assay robustness confirmed with Z' values between 0.7 and 0.8.

    Conclusions:

    • The developed cell-based assays are highly robust and well-suited for HTS.
    • These assays facilitate the discovery of drugs and probes targeting inflammatory pathways.
    • The study demonstrates the utility of phenotypic cell-based assays in inflammation research.