Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.

Détails

Ressource 1Télécharger: journal.pone.0192314.pdf (2936.74 [Ko])
Etat: Public
Version: Final published version
ID Serval
serval:BIB_5E840542134A
Type
Article: article d'un périodique ou d'un magazine.
Collection
Publications
Titre
Chromatin immunoprecipitation improvements for the processing of small frozen pieces of adipose tissue.
Périodique
PloS one
Auteur(s)
Castellano-Castillo D., Denechaud P.D., Moreno-Indias I., Tinahones F., Fajas L., Queipo-Ortuño M.I., Cardona F.
ISSN
1932-6203 (Electronic)
ISSN-L
1932-6203
Statut éditorial
Publié
Date de publication
2018
Peer-reviewed
Oui
Volume
13
Numéro
2
Pages
e0192314
Langue
anglais
Notes
Publication types: Journal Article ; Research Support, Non-U.S. Gov't
Publication Status: epublish
Résumé
Chromatin immunoprecipitation (ChIP) has gained importance to identify links between the genome and the proteome. Adipose tissue has emerged as an active tissue, which secretes a wide range of molecules that have been related to metabolic and obesity-related disorders, such as diabetes, cardiovascular failure, metabolic syndrome, or cancer. In turn, epigenetics has raised the importance in discerning the possible relationship between metabolic disorders, lifestyle and environment. However, ChIP application in human adipose tissue is limited by several factors, such as sample size, frozen sample availability, high lipid content and cellular composition of the tissue. Here, we optimize the standard protocol of ChIP for small pieces of frozen human adipose tissue. In addition, we test ChIP for the histone mark H3K4m3, which is related to active promoters, and validate the performance of the ChIP by analyzing gene promoters for factors usually studied in adipose tissue using qPCR. Our improvements result in a higher performance in chromatin shearing and DNA recovery of adipocytes from the tissue, which may be useful for ChIP-qPCR or ChIP-seq analysis.

Mots-clé
Adipose Tissue/metabolism, Adult, Chromatin Immunoprecipitation, Female, Freezing, Genome, Human, Humans, Male, Middle Aged, Proteome, Real-Time Polymerase Chain Reaction
Pubmed
Web of science
Open Access
Oui
Création de la notice
15/02/2018 17:25
Dernière modification de la notice
20/08/2019 14:16
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