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. 2017 Aug 23;7:9195. doi: 10.1038/s41598-017-09697-x

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© The Author(s) 2017

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Constitutive photo-activation of kRASG12V expression in localized tissue and single cells. To induce constitutive activation of kRASG12V, Cre-ERT2 mRNA was injected into Tg(ubi:loxP-Eos-stop-loxP-kRASG12V-T2A-mTFP) embryos at the one-cell stage. (A) Despite some minor light independent activation of Cre, mTFP expression could be fully induced by both cyclofen and caged cyclofen photo-activation. (B) Quantification of kRASG12V activation signal via the ratio of the fluorescence of the co-expressed mTFP protein and the (partially) floxed Eos protein. (C) UV (lamp or laser) and two-photon activation were performed at 10hpf to achieve global (with the UV lamp), local (with the UV laser) and single cell (with 2-photon laser) activation. Fish were imaged at 2dpf. Scale bar: 200 μm. Data are presented as mean ± SEM. ***p < 0.001; N.S, not significant.