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. 2018 Sep 10;8:13525. doi: 10.1038/s41598-018-31766-y

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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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Absence of NeuN and class III β-tubulin and presence of ubiquitin in CA. Representative images of human brain hippocampus sections immunostained with NeuN (a), anti-class III β-tubulin (TUJ1) (b) and anti-ubiquitin (Ubi) (c). As expected, when using isotype-specific anti-IgG antibodies conjugated to a red fluorochrome in the second incubation, NeuN stained the neuronal soma (arrowheads in a1), TUJ1 stains both neuronal soma and neurites (arrowheads in b1), and Ubi stained some dystrophic neurites (arrowheads in c1). CA were not labeled with NeuN (a1) and TUJ1 (yellow arrow in b2), but became labeled with Ubi (white arrow in c1). When using isotype-specific anti-IgM antibodies conjugated to a green fluorochrome in the second incubation, labeling of CA was observed in a2 and b3 (white arrows), but not in c2, indicating the presence of contaminant IgMs in the vials of NeuN and TUJ1, but not in the Ubi vial. Hoechst (blue) was used for nuclear staining. Scale bar: 50 μm.