Zelic Matija, Kelliher Michelle A
Department of Molecular, Cell and Cancer Biology, University of Massachusetts Medical School, Worcester, MA, USA.
Methods Mol Biol. 2018;1857:125-134. doi: 10.1007/978-1-4939-8754-2_12.
The serine/threonine kinase RIPK1 has numerous biological and pathological functions, mediating prosurvival as well as prodeath apoptotic and necroptotic signaling pathways downstream of various receptors, including death receptors and Toll-like receptors (TLRs). RIPK1 has been implicated in various diseases, including ischemia-reperfusion injury and inflammatory bowel disease (IBD). The recent generation of RIPK1 kinase inactive mice has enabled us to genetically interrogate the role of RIPK1 kinase-mediated necroptosis in disease models. Here, we describe procedures utilizing kinase inactive Ripk1 mice to analyze necroptosis induction in vitro in bone-marrow derived macrophages (BMDMs) and in vivo in a murine model of TNF-induced shock.
丝氨酸/苏氨酸激酶RIPK1具有多种生物学和病理学功能，介导多种受体（包括死亡受体和Toll样受体(TLRs)）下游的促生存以及促死亡的凋亡和坏死性凋亡信号通路。RIPK1与多种疾病有关，包括缺血再灌注损伤和炎症性肠病(IBD)。最近生成的RIPK1激酶失活小鼠使我们能够在疾病模型中通过基因手段探究RIPK1激酶介导的坏死性凋亡的作用。在此，我们描述了利用激酶失活的Ripk1小鼠分析骨髓来源的巨噬细胞(BMDM)体外坏死性凋亡诱导以及TNF诱导的休克小鼠模型体内坏死性凋亡诱导的实验步骤。
