Transcription Factors Downstream of IL-4 and TGF-β Signals: Analysis by Quantitative PCR, Western Blot, and Flow Cytometry

Atsushi Sugimoto; Ryoji Kawakami; Norihisa Mikami

doi:10.1007/978-1-4939-6877-0_11

Transcription Factors Downstream of IL-4 and TGF-β Signals: Analysis by Quantitative PCR, Western Blot, and Flow Cytometry

Methods Mol Biol. 2017:1585:141-153. doi: 10.1007/978-1-4939-6877-0_11.

Authors

Atsushi Sugimoto¹, Ryoji Kawakami¹, Norihisa Mikami²

Affiliations

¹ Department of Experimental Immunology, Immunology Frontier Research Center, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan.
² Department of Experimental Immunology, Immunology Frontier Research Center, Osaka University, 3-1 Yamadaoka, Suita, Osaka, 565-0871, Japan. mikami@ifrec.osaka-u.ac.jp.

PMID: 28477193
DOI: 10.1007/978-1-4939-6877-0_11

Abstract

IL-9-producing Th9 cell is a novel Th cell subset involved in type II allergic inflammations such as asthma. Th9 cells can be induced from naïve Th cells in the presence of IL-4 and TGF-β. It is also well established that downstream signals of IL-4 and TGF-β, including STAT6, IRF4, Smad, and PU.1, directly mediate IL-9 production in Th9 cells. In this chapter we describe the methods of flow cytometry, qPCR and western blot analysis to determine the expression or activation of these transcription factors downstream of IL-4 and TGF-β.

Keywords: IL-9; Intracellular staining; Th cell differentiation; Th9 cells.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Blotting, Western
Flow Cytometry
Humans
Interleukin-4 / metabolism*
Interleukin-9 / metabolism
Real-Time Polymerase Chain Reaction
T-Lymphocytes, Helper-Inducer / metabolism
Transforming Growth Factor beta / metabolism*

Substances

Interleukin-9
Transforming Growth Factor beta
Interleukin-4