Related Experiment Video

Updated: Jun 6, 2026

A Macrophage Reporter Cell Assay to Examine Toll-Like Receptor-Mediated NF-kB/AP-1 Signaling on Adsorbed Protein Layers on Polymeric Surfaces
07:55

A Macrophage Reporter Cell Assay to Examine Toll-Like Receptor-Mediated NF-kB/AP-1 Signaling on Adsorbed Protein Layers on Polymeric Surfaces

Published on: January 7, 2020

MicroRNA-101 targets MAPK phosphatase-1 to regulate the activation of MAPKs in macrophages

Qing-Yuan Zhu1, Qin Liu, Jian-Xia Chen

  • 1Institute Pasteur of Shanghai, Chinese Academy of Sciences, Shanghai, China.

Journal of Immunology (Baltimore, Md. : 1950)
|November 12, 2010
PubMed

Insights

MicroRNA-101 (miR-101) directly targets MAPK phosphatase-1 (MKP-1), regulating inflammatory responses. This microRNA influences macrophage activation by modulating MKP-1 expression, impacting innate immunity signaling pathways.

Area of Science:

  • Immunology
  • Molecular Biology
  • Genetics

Background:

  • MAPK phosphatase-1 (MKP-1) deactivates MAPKs and is implicated in regulating inflammatory cytokine biosynthesis.
  • The role of noncoding RNAs, specifically microRNAs, in MKP-1 expression and innate immune responses remains largely unexplored.

Purpose of the Study:

  • To investigate the role of microRNA-101 (miR-101) in regulating the expression of MAPK phosphatase-1 (MKP-1).
  • To elucidate the impact of miR-101 on macrophage innate immune responses, particularly in response to lipopolysaccharide (LPS).

Main Methods:

  • Dual-specificity phosphatase family analysis.
  • MicroRNA transfection and inhibition assays.
  • Western blotting to assess protein levels of MKP-1 and activated MAPKs (p38, JNK).
  • Toll-like receptor (TLR) ligand stimulation in RAW264.7 macrophage cells.
  • Inhibition of PI3K/Akt signaling pathway using LY294002 or Akt RNA interference.
  • Dexamethasone treatment effects on miR-101 and MKP-1 expression.

Main Results:

  • MKP-1 was identified as a direct target of miR-101.
  • miR-101 transfection attenuated LPS-induced MKP-1 expression and prolonged p38/JNK activation, while miR-101 inhibition enhanced MKP-1 and shortened MAPK activation.
  • Expression of miR-101 was induced by various TLR ligands (LPS, peptidoglycan, poly(I:C)).
  • PI3K/Akt pathway inhibition blocked LPS-induced miR-101 induction.
  • Dexamethasone inhibited miR-101 expression and enhanced MKP-1 in LPS-stimulated macrophages.

Conclusions:

  • miR-101 directly regulates MKP-1 expression, thereby modulating innate immune responses in macrophages.
  • The PI3K/Akt pathway is involved in the induction of miR-101 by LPS.
  • miR-101 serves as a key mediator in the inflammatory signaling cascade initiated by LPS in macrophages.

More Related Videos

Reprograming Model of Human Monocyte-derived Macrophages for In-vitro Assays08:37

Reprograming Model of Human Monocyte-derived Macrophages for In-vitro Assays

Published on: April 18, 2025

Related Concept Videos

MAPK Signaling Cascades01:07

MAPK Signaling Cascades

Mitogen-activated protein kinase, or MAPK pathway, activates three sequential kinases to regulate cellular responses such as proliferation, differentiation, survival, and apoptosis. The canonical MAPK pathway starts with a mitogen or growth factor binding to an RTK. The activated RTKs stimulate Ras, which recruits Raf or MAP3 Kinase (MAPKKK), the first kinase of the MAPK signaling cascade. Raf further phosphorylates and activates MEK or MAP2 Kinases (MAPKK), which in turn phosphorylates MAP...
PI3K/mTOR/AKT Signaling Pathway01:22

PI3K/mTOR/AKT Signaling Pathway

The mammalian target of rapamycin  (mTOR) is a serine/threonine kinase that regulates growth, proliferation, and cell survival in response to hormones, growth factors, or nutrient availability. This kinase exists in two structurally and functionally distinct forms: mTOR complex 1  (mTORC1) and mTOR complex 2  (mTORC2). The first form (mTORC1) is composed of a rapamycin-sensitive Raptor and proline-rich Akt substrate, PRAS40. In contrast,  mTORC2 consists of a rapamycin-insensitive companion...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns (non-coding regions of a gene) or intergenic regions (stretches of DNA present between genes). Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself, forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA...
MicroRNAs01:22

MicroRNAs

MicroRNA (miRNA) are short, regulatory RNA transcribed from introns—non-coding regions of a gene—or intergenic regions—stretches of DNA present between genes. Several processing steps are required to form biologically active, mature miRNA. The initial transcript, called primary miRNA (pri-mRNA), base-pairs with itself forming a stem-loop structure. Within the nucleus, an endonuclease enzyme, called Drosha, shortens the stem-loop structure into hairpin-shaped pre-miRNA. After the pre-miRNA ends...
mTOR Signaling and Cancer Progression03:03

mTOR Signaling and Cancer Progression

The mammalian target of rapamycin or mTOR protein was discovered in 1994 due to its direct interaction with rapamycin. The protein gets its name from a yeast homolog called TOR. The mTOR protein complex in mammalian cells plays a major role in balancing anabolic processes such as the synthesis of proteins, lipids, and nucleotides and catabolic processes, such as autophagy in response to environmental cues, such as availability of nutrients and growth factors.
The mTOR pathway or the...
The JAK-STAT Signaling Pathway01:20

The JAK-STAT Signaling Pathway

Several cytokine receptors have tightly bound Janus kinase or JAK proteins attached at their cytosolic tail. Small signaling molecules such as cytokines, growth hormones, or prolactins bind to the cytokine receptors and initiate their dimerization. The dimerization brings the cytosolic JAKs together that trans-phosphorylate and activates each other. The activated JAKs now phosphorylate cytosolic tails of the cytokine receptors, which serve as binding sites for adaptor proteins such as  SH2...