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 Concept Videos

You might also read

Related Articles

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

Sort by
Same author

Type I Interferon-Driven Monocyte Dysregulation and MAS-associated CD8 <sup>+</sup> T cells During Macrophage Activation Syndrome.

bioRxiv : the preprint server for biology·2026
Same author

A pathogenic gut lipoglycan drives systemic thromboinflammation in lupus nephritis.

Annals of the rheumatic diseases·2026
Same author

RPS19 and RPL5 haploinsufficient models reveal divergent ribosomal subunit controls of fetal hematopoiesis.

Nature communications·2026
Same author

Cyclin A2 induces cytokinesis in human adult cardiomyocytes and drives reprogramming in mice.

NPJ Regenerative medicine·2025
Same author

TLR-induced STK25 activation promotes IRF5-mediated inflammation.

Life science alliance·2025
Same author

Cyclin A2 Induces Human Adult Cardiomyocyte Cytokinesis and Elicits Cardiomyocyte Reprogramming and Dedifferentiation.

Research square·2025
Same journal

Editorial: Technologies for RNA Detection.

Bio-protocol·2026
Same journal

One-Step Affinity Purification of MarathonRT Reverse Transcriptase for RNA Sequencing Applications.

Bio-protocol·2026
Same journal

Enhanced RNA-Seq Expression Profiling and Functional Enrichment in Non-model Organisms Using Custom Annotations.

Bio-protocol·2026
Same journal

Using Combined Fluorescent In Situ Hybridization With Immunohistochemistry to Co-localize mRNA in Diverse Neuronal Cell Types.

Bio-protocol·2026
Same journal

Stepwise Protocol for Alternative Splicing Analysis in Single-Cell SMART-Seq2 RNA-Seq Data.

Bio-protocol·2026
Same journal

Enriching Bacteria-Specific RNA From Host Samples Before NGS With Transcript-Capture.

Bio-protocol·2026
See all related articles

Related Experiment Video

Updated: Nov 1, 2025

Native Polyacrylamide Gel Electrophoresis Immunoblot Analysis of Endogenous IRF5 Dimerization
08:57

Native Polyacrylamide Gel Electrophoresis Immunoblot Analysis of Endogenous IRF5 Dimerization

Published on: October 6, 2019

10.3K

Intracellular IRF5 Dimerization Assay.

Cherrie D Sherman1, Betsy J Barnes1

  • 1Center for Autoimmune Musculoskeletal and Hematopoietic Diseases, Feinstein Institutes for Medical Research, Manhasset, New York, USA.

Bio-Protocol
|June 21, 2021
PubMed
Summary
This summary is machine-generated.

This study introduces two assays to detect interferon regulatory factor 5 (IRF5) homodimerization and interactions with cell-penetrating peptide (CPP) inhibitors. These methods offer rapid and accurate detection of IRF5 molecular interactions in cells.

Keywords:
DimerizationFRETIRF5IRF5-CPPMolecular interactionNative-PAGEPolyacrylamide gel electrophoresis

More Related Videos

Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice
08:09

Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice

Published on: March 24, 2017

8.3K
A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3
11:44

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

12.2K

Related Experiment Videos

Last Updated: Nov 1, 2025

Native Polyacrylamide Gel Electrophoresis Immunoblot Analysis of Endogenous IRF5 Dimerization
08:57

Native Polyacrylamide Gel Electrophoresis Immunoblot Analysis of Endogenous IRF5 Dimerization

Published on: October 6, 2019

10.3K
Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice
08:09

Vasodilation of Isolated Vessels and the Isolation of the Extracellular Matrix of Tight-skin Mice

Published on: March 24, 2017

8.3K
A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3
11:44

A High Resolution Method to Monitor Phosphorylation-dependent Activation of IRF3

Published on: January 24, 2016

12.2K

Area of Science:

  • Immunology
  • Molecular Biology
  • Biochemistry

Background:

  • Interferon regulatory factor 5 (IRF5) plays a crucial role in innate immunity and inflammatory responses.
  • Understanding IRF5's molecular interactions, such as homodimerization and binding to inhibitors, is vital for deciphering its regulatory mechanisms.
  • Existing methods for studying endogenous IRF5 interactions are limited.

Purpose of the Study:

  • To develop and validate novel assays for detecting endogenous IRF5 homodimerization.
  • To establish a method for assessing the interaction between endogenous IRF5 and cell-penetrating peptide (CPP) inhibitors.
  • To provide tools for rapid and accurate analysis of IRF5 molecular interactions.

Main Methods:

  • Native-polyacrylamide gel electrophoresis (PAGE) followed by immunoblotting to detect IRF5 homodimers in THP-1 cells treated with an IRF5-targeted CPP (IRF5-CPP) inhibitor.
  • In-cell fluorescence resonance energy transfer (FRET) assay to measure the interaction between endogenous IRF5 and FITC-labeled IRF5-CPP in THP-1 cells.
  • Stimulation of THP-1 cells with R848 to activate IRF5 signaling pathways.

Main Results:

  • Successfully detected endogenous IRF5 homodimers using native-PAGE and immunoblotting.
  • Demonstrated the ability to measure IRF5-CPP interactions using an in-cell FRET assay.
  • Quantified fluorescence transfer as FRET units, indicating successful molecular interaction detection.

Conclusions:

  • The developed native-PAGE and FRET assays are effective for detecting endogenous IRF5 homodimerization and IRF5-CPP interactions.
  • These assays provide rapid, accurate, and robust methods for studying IRF5 molecular interactions.
  • The findings facilitate further research into IRF5 function and the development of targeted therapeutics.