Department of Genetics, Graduate School of Medicine, Osaka University, Suita, Osaka 565-0871, Japan.
Research Institute for Microbial Diseases (RIMD), Osaka University, Suita, Osaka 565-0871, Japan; Immunology Frontier Research Center (IFReC), Osaka University, Suita, Osaka 565-0871, Japan.
STAR Protoc. 2022 Sep 16;3(3):101539. doi: 10.1016/j.xpro.2022.101539. Epub 2022 Jul 11.
Senescent cells accumulation is associated with aging and age-related diseases, and recent findings suggest that autophagy, the activity of the intracellular degradation system, decreases during senescence. In this protocol, we detail steps to induce cellular senescence in response to DNA damage, evaluate the senescent state using SA-β-gal staining and western blot for p21, LAMP1, and Lamin B1, and detect autophagy via LC3 western blotting. This protocol can be used in most cell lines and for various types of senescent cells. For complete details on the use and execution of this protocol, please refer to Yamamoto-Imoto et al. (2022).
衰老细胞的积累与衰老和与年龄相关的疾病有关，最近的研究结果表明，自噬——细胞内降解系统的活性——在衰老过程中会下降。在本方案中，我们详细介绍了如何在 DNA 损伤的情况下诱导细胞衰老，使用 SA-β-半乳糖苷染色和 p21、LAMP1 和 Lamin B1 的 Western blot 来评估衰老状态，以及通过 LC3 Western blot 检测自噬。本方案可用于大多数细胞系和各种类型的衰老细胞。有关该方案使用和执行的完整详细信息，请参阅 Yamamoto-Imoto 等人。(2022)。
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