Analysis of suppressor of cytokine signalling (SOCS) gene expression by real-time quantitative PCR

Abstract

The Suppressor of Cytokine Signalling (SOCS) proteins are a family of negative regulators characterized by a central SH2 domain and C-terminal SOCS box motif. Cytokine Inducible SH2-containing protein (CIS), SOCS1, 2 and 3 are rapidly upregulated in response to cytokine stimulation and act to inhibit JAK/ STAT signalling by a variety of mechanisms. The expression of SOCS proteins provides a level of specificity in the control of signalling, with SOCS proteins differentially upregulated in response to individual cytokines and in various cell-types. Real-time reverse transcription (RT) quantitative polymerase chain reaction (RT-qPCR) is an established technique for quantifying mRNA in biological samples, measuring the relative expression of genes of interest and identifying single nucleotide polymorphisms. Here we describe the use of SYBR ® Green I RT-qPCR to quantify the relative expression level of SOCS mRNA in murine bone marrow-derived macrophages (BMDM). The approach can be universally applied to different cell types and various tissues.