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 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)