Transcriptional regulation in adipogenesis through PPARγ-dependent and -independent mechanisms by prostaglandins

Methods Mol Biol. 2014;1164:177-96. doi: 10.1007/978-1-4939-0805-9_15.

Abstract

Adipogenesis is controlled by complex mechanisms, and transcription factors are involved in its regulation. PPARγ is a ligand-dependent transcription factor and the most important one for adipogenesis. Although prostaglandin (PG) D2 metabolites have been reported as being the ligands of PPARγ, the endogenous PPARγ ligand in adipocytes remains unclear. Here, we show the methods for the general analysis of adipocyte differentiation and the protocols for promoter analysis, fluorescence EMSA, and chromatin immunoprecipitation assay for the transcriptional regulation of the SREBP-1c-activated lipocalin-type PGD synthase gene in adipocytes. Moreover, we describe that PGD2 and its metabolites are involved in the regulation of adipogenesis through PPARγ-dependent and -independent mechanisms.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • 3T3-L1 Cells
  • Adipocytes / cytology*
  • Adipocytes / metabolism
  • Adipogenesis*
  • Animals
  • Azo Compounds / analysis
  • Chromatin Immunoprecipitation / methods
  • Coloring Agents / analysis
  • Fluorescence
  • Gene Expression Regulation, Developmental
  • Genetic Vectors / genetics
  • Intramolecular Oxidoreductases / genetics*
  • Lipocalins / genetics*
  • Mass Spectrometry / methods
  • Mice
  • PPAR gamma / metabolism*
  • Plasmids / genetics
  • Promoter Regions, Genetic
  • Prostaglandins / metabolism*
  • RNA, Messenger / analysis
  • RNA, Messenger / genetics
  • Staining and Labeling / methods
  • Transcriptional Activation*
  • Two-Hybrid System Techniques

Substances

  • Azo Compounds
  • Coloring Agents
  • Lipocalins
  • PPAR gamma
  • Prostaglandins
  • RNA, Messenger
  • Intramolecular Oxidoreductases
  • prostaglandin R2 D-isomerase
  • oil red O