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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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Tumor induced by periodic or constitutive, but not transient activation of kRASG12V expression. In the latter system, one-time activation of kRASG12V expression was induced by transient cyclofen incubation or photo-activation of caged cyclofen at 1dpf (A). Periodic kRASG12V expression was achieved by 1 day incubation in 2 μM cyclofen every 5 days for a course of two months (E). (B–D) Transient induction of kRASG12V did not affect normal development (as shown by the histopathological staining) with no CFP expression in one-year old fish. (E–H) Tumors were observed (albeit rarely) in fish periodically treated with cyclofen. A representative one-year old fish displayed a clear tumor (E) revealed by histopathological staining (H). Interestingly, the cells in the tumor still expressed EosFP (F), but lost CFP expression (G). (I–L) Constitutive kRASG12V expression following light-activation of caged cyclofen induced tumor (I) in 5 month-old fish as revealed by both fluorescent markers (J,K) and H&E staining (L). Scale bar: B, D, F, H, L 200 μm; J, 1 cm.