Cardiovascular Division, Washington University School of Medicine, St Louis, MO, USA.
Division of Endocrinology, Metabolism, and Lipid Research, Washington University School of Medicine, St Louis, MO, USA.
STAR Protoc. 2022 Dec 16;3(4):101665. doi: 10.1016/j.xpro.2022.101665. Epub 2022 Sep 11.
Previous studies have demonstrated that a high-protein diet leads to increased atherosclerosis in mice, and that this adverse effect is caused by activation of macrophage mTORC1 signaling. Here, we provide a detailed protocol for the evaluation of diet-induced mTORC1 signaling in plaque macrophages in atherosclerosis-prone apolipoprotein E (ApoE) knockout (KO) mice. This protocol includes flow cytometry and immunofluorescence analysis of atherosclerotic macrophages that can be used to study the atherogenic potential of a variety of mTORC1 modulators. For complete details on the use and execution of this protocol, please refer to Zhang et al. (2020).
先前的研究表明，高蛋白饮食会导致小鼠动脉粥样硬化加剧，而这种不利影响是由巨噬细胞 mTORC1 信号的激活引起的。在这里，我们提供了一种详细的方案，用于评估易发生动脉粥样硬化的载脂蛋白 E（ApoE）基因敲除（KO）小鼠中斑块巨噬细胞中饮食诱导的 mTORC1 信号。该方案包括对动脉粥样硬化巨噬细胞进行流式细胞术和免疫荧光分析，可用于研究各种 mTORC1 调节剂的动脉粥样硬化形成潜力。有关此方案的使用和执行的完整详细信息，请参阅 Zhang 等人（2020 年）。
