使用流式细胞术和蛋白质印迹法分析葡聚糖硫酸钠诱导的结肠炎小鼠中转化生长因子β信号传导的实验方案。
Protocol for analyzing transforming growth factor β signaling in dextran-sulfate-sodium-induced colitic mice using flow cytometry and western blotting.
作者信息
Fujiwara Mai, Garo Lucien, Ajay Amrendra K, Cannon Alkeiver S, Kolypetri Panagiota, Dhuppar Shivnarayan, Murugaiyan Gopal
机构信息
Ann Romney Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA.
Ann Romney Center for Neurologic Diseases, Department of Neurology, Brigham and Women's Hospital and Harvard Medical School, Boston, MA 02115, USA; Boston University School of Medicine, Boston, MA 02118, USA.
出版信息
STAR Protoc. 2023 Apr 25;4(2):102249. doi: 10.1016/j.xpro.2023.102249.
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..
转化生长因子β（TGF-β）对于维持肠道免疫稳态至关重要。在此，我们展示了在葡聚糖硫酸钠诱导的结肠炎小鼠中分析TGF-β受体信号下游Smad分子的技术。我们描述了结肠炎的诱导、细胞分离以及树突状细胞和T细胞的流式细胞术细胞分选。然后，我们详细介绍了磷酸化Smad2/3的细胞内染色以及Smad7的蛋白质印迹分析。该方案可在来自许多来源的有限数量的细胞上进行。有关此方案的使用和执行的完整详细信息，请参考加罗等人的研究。
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