Departamento de Bioquímica-Instituto de Investigaciones Biomédicas Sols-Morreale (IIBM), Universidad Autónoma de Madrid-Consejo Superior de Investigaciones Científicas, Madrid, Spain.
Methods Mol Biol. 2025;2876:77-91. doi: 10.1007/978-1-0716-4252-8_5.
Apoptosis is a type of programmed cell death which can be induced by thyroid hormone in pituitary and other cell models. This process is characterized by several biochemical changes, including modifications in plasma membrane lipid composition, mitochondrial membrane dysfunction, and DNA fragmentation, among others. These cellular alterations can be measured using western blotting and flow cytometry techniques. Here we describe the method to detect cleavage of poly-ADP ribose polymerase by western blotting, as well as different flow cytometry-based methods to quantify the sub-G1 hypodiploid cell population, the redistribution of phosphatidylserine to the outer leaflet of plasma membrane, and the loss of the mitochondrial inner membrane potential.
细胞凋亡是一种由甲状腺激素诱导的细胞程序性死亡，它可以发生在垂体和其他细胞模型中。这个过程的特点是发生了几种生化变化，包括质膜脂质组成的改变、线粒体膜功能障碍和 DNA 片段化等。这些细胞变化可以通过 Western blot 和流式细胞术技术来测量。在这里，我们描述了通过 Western blot 检测聚 ADP 核糖聚合酶裂解的方法，以及基于流式细胞术的几种不同方法，用于定量检测亚 G1 低倍体细胞群、磷脂酰丝氨酸向质膜外层的重新分布以及线粒体膜电位的丧失。
