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. 2019 Mar 5;9:3545. doi: 10.1038/s41598-019-40310-5

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© The Author(s) 2019

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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d16HER2 and WTHER2 isoform detection in HER2-positive BC (ZR75.30), GC (N87) and Oesophago-gastric (OGC) (OE19) and HER2-negative GC (MKN45) cell lines evaluated using qPCR, immunohistochemistry (IHC) and mRNA bright-field in situ hybridization (ISH) analyses. (A) qPCR analysis of relative d16HER2 and WTHER2 expression; (B–Q) HER2 expression by IHC and WTHER2 and d16HER2 isoforms detection by mRNA ISH. The ZR75.30 cell line (B–E) exhibits positive HER2 IHC reactivity (B); CRHER2 and WTHER2 mRNA ISH (score 4) (C,D); d16HER2 mRNA ISH (score 3) (E). The N87 cell line (F,G,H,I) exhibits positive HER2 IHC reactivity (F); CRHER2 and WTHER2 mRNA ISH (score 4) (G,H); d16HER2 mRNA ISH (score 2) (I). The OE19 cell line (J–M) exhibits positive HER2 IHC reactivity (J); CRHER2 and WTHER2 mRNA ISH (score 4) (K,L); d16HER2 mRNA ISH (score 3) (M). The MKN45 cell line (N,O,P,Q) exhibits negative HER2 IHC reactivity (N); CRHER2 and WTHER2 mRNA ISH (score 1) (O,P); d16HER2 mRNA ISH (score 0) (Q). Original magnification: 20 × (B,F,J,N; scale bar: 25 μm) and 40 × (C–Q; scale bar: 12,5 μm).