Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
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Updated: Jul 9, 2025
Mapping the Cellular Distribution of an Optogenetic Protein Using a Light-Stimulation Grid
Published on: January 26, 2024
Yoshibumi Ueda1, Shohei Matsushita2, Mitsugu Suzuki3
1Department 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.
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.
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