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. 2018 Mar 6;8:4070. doi: 10.1038/s41598-018-22434-2

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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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Representative in situ RT-PCR/ FISH for IR/A and IR/B along with MAP2 immunohistochemistry in the frontal cortex of the human brain. (A) FFPE frontal cortex human brain tissue (7 µm) was assayed by RT-PCR/FISH for (B) IR/A (green) and (C) IR/B (red) followed by immunohistochemistry for the neuronal marker (D) MAP2. Sections were mounted with Vectashield containing (E) DAPI (blue) to visualize the nuclei. Breakout of (A) shows the co-localization of the red (IR/A) and green (IR/B) signals and specifically a neuron expressing only the IR/A isoform (yellow arrow). (F) Staining analysis of IR/A (red) and IR/B (green) signal by digital fluorescent microscopy and Squassh. (G) Co-localization analysis of IR/A (green) and IR/B (red) signal with MAP2 (DAB) immunohistochemistry. White arrow indicates co-localization of IR/A, IR/B and MAP2 staining. Yellow arrow indicates co-localization of IR/B only with MAP2 staining. *Indicates statistical significance compared to IR/A or IR/B alone (p < 0.01) using ANOVA with Tukey’s multiple comparison. Scale bar = 25 µm.