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

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...

Descripción completa

Detalles Bibliográficos
Autores principales: Ueda, Yoshibumi, Matsushita, Shohei, Suzuki, Mitsugu, Ozawa, Takeaki
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10628809/
http://dx.doi.org/10.1016/j.xpro.2023.102622
_version_ 1785131840339705856
author Ueda, Yoshibumi
Matsushita, Shohei
Suzuki, Mitsugu
Ozawa, Takeaki
author_facet Ueda, Yoshibumi
Matsushita, Shohei
Suzuki, Mitsugu
Ozawa, Takeaki
author_sort Ueda, Yoshibumi
collection PubMed
description 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)
format Online
Article
Text
id pubmed-10628809
institution National Center for Biotechnology Information
language English
publishDate 2023
publisher Elsevier
record_format MEDLINE/PubMed
spelling pubmed-106288092023-11-08 Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns Ueda, Yoshibumi Matsushita, Shohei Suzuki, Mitsugu Ozawa, Takeaki STAR Protoc Protocol 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) Elsevier 2023-10-27 /pmc/articles/PMC10628809/ http://dx.doi.org/10.1016/j.xpro.2023.102622 Text en © 2023 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Ueda, Yoshibumi
Matsushita, Shohei
Suzuki, Mitsugu
Ozawa, Takeaki
Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
title Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
title_full Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
title_fullStr Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
title_full_unstemmed Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
title_short Protocol for screening cellular outputs activated by optogenetically controlled temporal PI3K signaling activation patterns
title_sort protocol for screening cellular outputs activated by optogenetically controlled temporal pi3k signaling activation patterns
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10628809/
http://dx.doi.org/10.1016/j.xpro.2023.102622
work_keys_str_mv AT uedayoshibumi protocolforscreeningcellularoutputsactivatedbyoptogeneticallycontrolledtemporalpi3ksignalingactivationpatterns
AT matsushitashohei protocolforscreeningcellularoutputsactivatedbyoptogeneticallycontrolledtemporalpi3ksignalingactivationpatterns
AT suzukimitsugu protocolforscreeningcellularoutputsactivatedbyoptogeneticallycontrolledtemporalpi3ksignalingactivationpatterns
AT ozawatakeaki protocolforscreeningcellularoutputsactivatedbyoptogeneticallycontrolledtemporalpi3ksignalingactivationpatterns

Ejemplares similares