Related Experiment Video

Updated: Aug 20, 2026

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization
13:55

Combined Immunofluorescence and DNA FISH on 3D-preserved Interphase Nuclei to Study Changes in 3D Nuclear Organization

Published on: February 3, 2013

Reductions in I kappa B epsilon and changes in NF-kappa B activity during B lymphocyte differentiation

Stefan Doerre1, Kristin Perkins Mesires, Kylle M Daley

  • 1Department of Microbiology, Boston University School of Medicine, Boston, MA 02118, USA.

Journal of Immunology (Baltimore, Md. : 1950)
|January 7, 2005
PubMed

Insights

IkappaBepsilon protein levels decrease upon B cell stimulation, transiently altering NF-kappaB activity. This suggests IkappaBepsilon regulates nuclear NF-kappaB, impacting B cell differentiation.

Area of Science:

  • Immunology
  • Molecular Biology
  • Cell Signaling

Background:

  • The NF-kappaB signaling pathway is crucial for immune responses.
  • IkappaB proteins regulate NF-kappaB activity by sequestering it in the cytoplasm.
  • Specific roles of different IkappaB family members, like IkappaBepsilon, in B cell activation are not fully understood.

Purpose of the Study:

  • To investigate the expression and stability of IkappaBepsilon in splenic B cells.
  • To compare IkappaBepsilon regulation with IkappaBalpha and IkappaBbeta during B cell stimulation.
  • To elucidate the role of IkappaBepsilon in modulating NF-kappaB activity and gene expression in B cells.

Main Methods:

  • Analysis of IkappaBepsilon, IkappaBalpha, and IkappaBbeta protein levels in murine splenic B cells.
  • Stimulation of B cells in vitro and in vivo using various agents.
  • Assessment of NF-kappaB reporter construct activity sensitive to p65/RelA homodimers.
  • Comparison of NF-kappaB-responsive gene expression in IgM(+) and IgG(+) B cell lines.

Main Results:

  • Primary splenic B cells express high levels of IkappaBepsilon, comparable to IkappaBalpha.
  • Unlike IkappaBalpha and IkappaBbeta, IkappaBepsilon is rapidly degraded upon B cell stimulation.
  • B cell stimulation leads to a transient increase in NF-kappaB activity, correlating with IkappaBepsilon reduction.
  • Differences in NF-kappaB-responsive gene expression exist between IgM(+) and IgG(+) B cell subsets.

Conclusions:

  • IkappaBepsilon plays a significant role in the transient regulation of nuclear NF-kappaB activity in stimulated B cells.
  • Reductions in IkappaBepsilon levels contribute to dynamic changes in NF-kappaB activity during B cell activation.
  • These transient NF-kappaB activity changes may influence long-term alterations associated with B cell differentiation.

More Related Videos

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α11:27

A Guide to Production, Crystallization, and Structure Determination of Human IKK1/α

Published on: November 2, 2018

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells10:57

NF-κB-dependent Luciferase Activation and Quantification of Gene Expression in Salmonella Infected Tissue Culture Cells

Published on: January 12, 2020

Related Concept Videos

NF-κB-dependent Signaling Pathway02:26

NF-κB-dependent Signaling Pathway

The transcription factor NF-κB was discovered in 1986 in the lab of Nobel laureate Professor David Baltimore, for its interaction with the immunoglobulin light chain enhancer in B-cells. After more than three decades of study, it is now evident that NF-κB regulates the expression of over 100 genes. Most of these genes play an essential role in the innate and adaptive immune responses as well as the inflammatory responses of animals.
NF-κB-dependent Signaling Mechanism
The heterodimer of NF-κB...
Co-activators and Co-repressors02:04

Co-activators and Co-repressors

Gene transcription is regulated by the synergistic action of several proteins that form a complex at a gene regulatory site. This is observed in eukaryotes, where the regulation of gene expression is a complex process. Regulatory proteins in eukaryotes can broadly be classified into two types – regulators that bind directly to specific DNA sequences and co-regulators that associate with regulatory proteins but cannot directly bind to the DNA. These co-regulators are further divided into...
Receptor Downregulation in MVBs01:15

Receptor Downregulation in MVBs

Multivesicular bodies (MVBs) are mature endosomes that sort ubiquitinated proteins and then fuse with lysosomes to degrade the sorted proteins. Epidermal growth factor (EGF) and its receptor (EGFR) form a complex that can be internalized through endocytosis, sorted into an MVB, and later degraded.
The EGFR can initiate signaling pathways that  lead to cell proliferation, migration, and differentiation. Overexpression of EGFR  stimulates cells to proliferate. Excessive  EGFR activation may...
The Intrinsic Apoptotic Pathway01:31

The Intrinsic Apoptotic Pathway

Internal cellular stress, such as cellular injury or hypoxia, triggers intrinsic apoptosis. The B-cell lymphoma 2 (Bcl-2) family of proteins are the primary regulators of the intrinsic apoptotic pathway. For example, during DNA damage, checkpoint proteins, such as Ataxia Telangiectasia Mutated (ATM protein) and Checkpoints Factor-2 (Chk2) proteins, are activated. These proteins phosphorylate p53 which further activates pro-apoptotic proteins, such as Bax, Bak, PUMA, and Noxa, and inhibits...
Abnormal Proliferation02:23

Abnormal Proliferation

Under normal conditions, most adult cells remain in a non-proliferative state unless stimulated by internal or external factors to replace lost cells. Abnormal cell proliferation is a condition in which the cell's growth exceeds and is uncoordinated with normal cells. In such situations, cell division persists in the same excessive manner even after cessation of the stimuli, leading to persistent tumors. The tumor arises from the damaged cells that replicate to pass the damage to the daughter...
The Nucleolus02:55

The Nucleolus

The nucleolus is the most prominent substructure of the nucleus. When it was first discovered, it was considered to be an isolated organelle that forms fibrils and granules. In 1931, the relationship between the nucleolus and chromosomes was first described by Heitz. He observed that the appearance and size of nucleolus varies depending on the stage of the cell cycle. He also noticed constricted regions on different chromosomes clustered together at definite cell cycle stages. These regions,...