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. 2018 Sep 10;8:13545. doi: 10.1038/s41598-018-31787-7

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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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Lunasin can block cell death to rescue Aβ42 mediated neurodegeneration. The dying cells nuclei can be marked by TUNEL staining. TUNEL staining was carried out in (A,A’) Wild-type, (B,B’) GMR > Aβ42, (C,C’) GMR > Aβ42 + Lun eye imaginal discs. The number of dying retinal neurons were counted in these backgrounds (n = 5). Note that the number of dying cells increase nearly 3–4 fold in (B,D) GMR > Aβ42 as compared to the (A,D) Wild-type eye discs. (C,D) Misexpression of Lunasin (Lun) along with GMR > Aβ42 (GMR > Aβ42 + Lun-GFP) results in significant reduction in the dying retinal neurons. The number of TUNEL positive nuclei were counted from five eye imaginal discs for all three backgrounds. (D) A graph comparing the number of dying nuclei of neurons validate that Lun misexpression along with GMR > Aβ42 (GMR > Aβ42 + Lun-GFP) rescues the GMR > Aβ42 neurodegeneration. These numbers are significant based on the calculations of P-values using the two-tailed t- test using Microsoft Excel 2013. (E–H) Accumulation of amyloid plaque was detected using monoclonal antibody 6E10 in (E) Wild-type, (F) GMR > Aβ42 and (G) GMR > Aβ42 + Lun eye imaginal discs. (H) The signal intensity of 6E10 staining was calculated from five (n = 5) eye discs of each background and plotted on a graph. Note that 6E10 levels are not significantly different between (F) GMR > Aβ42 and (G) GMR > Aβ42 + Lun background. The levels of amyloid plaques are barely detected in Wild-type background. Magnification of all eye disc is 20X.