title: 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.
authors.affiliation: Inflammation Division, Walter and Eliza Hall Institute of Medical Research, Parkville, VIC, Australia. kolesnik@wehi.edu.au
authors.forename: Tatiana B
authors.initials: TB
authors.lastname: Kolesnik
authors.forename: Sandra E
authors.initials: SE
authors.lastname: Nicholson
mesh_terms: D001854:Bone Marrow Cells / Q000166:cytology / Q000187:drug effects; D017931:DNA Primers / Q000235:genetics; D018076:DNA, Complementary / Q000096:biosynthesis; D005786:Gene Expression Regulation; D006801:Humans; D015847:Interleukin-4 / Q000494:pharmacology; D008264:Macrophages / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction / Q000379:methods* / Q000592:standards; D012015:Reference Standards; D050826:Suppressor of Cytokine Signaling Proteins / Q000235:genetics*
chemical_list: D017931:DNA Primers; D018076:DNA, Complementary; D050826:Suppressor of Cytokine Signaling Proteins; D015847:Interleukin-4