Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns

Yoshibumi Ueda; Shohei Matsushita; Mitsugu Suzuki; Takeaki Ozawa

doi:10.1016/j.xpro.2023.102622

Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns

STAR Protoc. 2023 Dec 15;4(4):102622. doi: 10.1016/j.xpro.2023.102622. Epub 2023 Oct 27.

Authors

Yoshibumi Ueda¹, Shohei Matsushita², Mitsugu Suzuki³, Takeaki Ozawa⁴

Affiliations

¹ Department of Chemistry, School of Science, The University of Tokyo, Tokyo, Japan; Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, Tokyo, Japan. Electronic address: yoshibumiueda@gmail.com.
² Department of Chemistry, School of Science, The University of Tokyo, Tokyo, Japan.
³ Department of Quality Assurance and Radiological Protection, National Institute of Infectious Diseases, Tokyo, Japan.
⁴ Department of Chemistry, School of Science, The University of Tokyo, Tokyo, Japan. Electronic address: ozawa@chem.s.u-tokyo.ac.jp.

Abstract

PI3K signaling elicits distinct outputs in response to different patterns of extracellular stimulation. Here, we present a protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns in 96-well plates. We describe steps for establishing PPAP2-stable cells, probe expression, and blue light irradiation. We then detail procedures for analysis of translation activity. This protocol can be applied for purposes, such as examining the effect of PI3K signaling on the efficacy of anticancer drugs. For complete details on the use and execution of this protocol, please refer to Ueda et al. (2022).¹.

Keywords: Cancer; Cell Biology; Cell Membrane.

MeSH terms

Blue Light*
Phosphatidylinositol 3-Kinases*

Substances

Phosphatidylinositol 3-Kinases