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. 2018 May 4;8:7072. doi: 10.1038/s41598-018-25328-5

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© The Author(s) 2018

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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Multi-plex in situ mutation detection of MET using padlock probe rolling circle amplification on E98-FM FFPE xenograft tissue. (a) Serial H&E stainings. (b) In situ duo-probe detection of METexon7–8^wt and MET^Δ7-8 transcripts using padlock probes MET^7,8 and MET^6–9, respectively. (c) In situ duo-probe detection of METexon14^wt and MET^Δ14 transcripts using padlock probes MET¹⁴ and MET^13–15, respectively. (d) In situ duo-probe detection of METexon14^wt and MET^Δ7-8 transcripts using padlock probes MET¹⁴ and MET^6–9, respectively. Wild-type transcripts are displayed as green RCPs, mutant transcripts are displayed as magenta RCPs. Cell nuclei are shown in grey. Note that in all assays the tumour border emphasizes the specificity of the padlock probes, showing no fluorescent detection spots in normal brain tissue (right figures, original magnification 20x). Compare the left and middle panels for each assay, visualizing differences in transcript expression in different tumour areas. Inserts give a zoomed view of the underlying image. Original magnification 40x.