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. Author manuscript; available in PMC: 2022 Jul 4.
Published in final edited form as: Nat Chem Biol. 2021 Dec 22;18(2):152–160. doi: 10.1038/s41589-021-00917-0

Figure 1. Single-component BcLOV4 fusions allow control of Ras and PI3K signaling.

Figure 1.

A) BcLOV4 binds the cell membrane when exposed to blue light. The three BcLOV4 domains represent the LOV, DUF, and RGS domains, as previously described15 B) Light-induced membrane recruitment of BcLOV4 fused to the SOScat catalytic domain will induce Ras/Erk signaling. C) Analogous recruitment of the iSH domain will induce PI3K/Akt signaling. D,E) Five minutes of blue light stimulation (160 mW/cm2 at 20% duty cycle) increases intracellular ppErk levels in cells that express BcLOV-SOScat (D) and increases pAkt in cells that express BcLOV-iSH (E). Grey zone indicates the change in ppErk or pAkt in wild-type cells that were stimulated with 10% calf serum for 10 minutes. Data represent means of three biologically independent replicates, each representing the mean signal intensity from ~2000-4000 single cells. F,G) Light intensity dose-response of (F) ppErk fold-change induction in BcLOV-SOScat-expressing cells or (G) pAkt fold-change induction in BcLOV-iSH cells at 100% duty cycle after 5 minutes of illumination. Data represent means +/− SD of three biologicially independent replicates, each representing the mean signal intensity from ~300-500 single cells. All stimulation in A-G was achieved using the optoPlate-96. All stimulation and environmental conditions for all figures can be found in Supplementary Table 1.