Protocol for analyzing transforming growth factor β signaling in dextran-sulfate-sodium-induced colitic mice using flow cytometry and western blotting

使用流式细胞术和蛋白质印迹法分析葡聚糖硫酸钠诱发的结肠炎小鼠中转化生长因子 β 信号传导的方案

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作者:Mai Fujiwara, Lucien Garo, Amrendra K Ajay, Alkeiver S Cannon, Panagiota Kolypetri, Shivnarayan Dhuppar, Gopal Murugaiyan

Abstract

Transforming growth factor β (TGF-β) is critical to the maintenance of intestinal immune homeostasis. Here, we present techniques for analyzing Smad molecules downstream of TGF-β receptor signaling in dextran-sulfate-sodium-induced colitic mice. We describe colitis induction, cell isolation, and flow cytometric cell sorting of dendritic cells and T cells. We then detail intracellular staining of phosphorylated Smad2/3 and western blotting analysis of Smad7. This protocol can be performed on a limited number of cells from many sources. For complete details on the use and execution of this protocol, please refer to Garo et al.1.

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