Protocol for photoactivation of YAP in cancer cell spheroids embedded in collagen gels

STAR Protoc. 2021 Jul 7;2(3):100657. doi: 10.1016/j.xpro.2021.100657. eCollection 2021 Sep 17.

Abstract

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).

Keywords: Antibody; Biotechnology and bioengineering; Cancer; Cell Biology; Cell culture; Microscopy; Molecular/Chemical Probes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Culture Techniques / methods*
  • Collagen / chemistry
  • Fluorescent Antibody Technique
  • Gels / chemistry
  • HeLa Cells
  • Humans
  • Optogenetics / methods*
  • Plasmids / genetics
  • Spheroids, Cellular* / cytology
  • Spheroids, Cellular* / metabolism
  • Tumor Cells, Cultured* / cytology
  • Tumor Cells, Cultured* / metabolism
  • YAP-Signaling Proteins* / genetics
  • YAP-Signaling Proteins* / metabolism

Substances

  • Gels
  • YAP-Signaling Proteins
  • Collagen