Department of Chemistry and Center for NanoScience (CeNS), Ludwig-Maximilians-Universität München, Butenandtstr. 11, 81377 Munich, Germany.
Institute of Pharmaceutical Sciences, Department of Pharmaceutical Chemistry, University of Graz, Humboldtstr. 46, 8010 Graz, Austria.
STAR Protoc. 2021 Jul 7;2(3):100657. doi: 10.1016/j.xpro.2021.100657. eCollection 2021 Sep 17.
This protocol describes the necessary preparations and procedures to photo-activate Yes-associated protein (YAP) with optoYAP in cancer cell spheroids in 3D collagen matrices. We detail steps for immunofluorescent staining of the resulting YAP-activated HeLa spheroids. In addition, we describe handling of optoYAP on 2D substrates. While this protocol focuses on the use of optoYAP in 3D HeLa cell culture, it can be modified for other cell types. For complete details on the use and execution of this protocol, please refer to Illes et al. (2021).
本方案描述了在 3D 胶原基质中的癌细胞球体中用光激活 YAP 的必要准备和步骤。我们详细介绍了对由此产生的 YAP 激活的 HeLa 球体进行免疫荧光染色的步骤。此外，我们还描述了在 2D 基质上处理 optoYAP 的方法。虽然本方案专注于在 3D HeLa 细胞培养中使用 optoYAP，但它可以针对其他细胞类型进行修改。有关使用和执行本方案的完整详细信息，请参阅 Illes 等人（2021 年）。
