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. 2019 Aug 13;9:11782. doi: 10.1038/s41598-019-48279-x

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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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(a) Effects of furin inhibitor on the levels of α-chain, β-chain, and ICD of LRP1 at 4 h after 0 µM (0) or 30 µM (30) NMDA treatment with (+) or without (−) 10 µM furin inhibitor. Proteins from cortical neurons in primary culture were analyzed by Western blotting with anti-α-chain and anti-β-chain of LRP1/ICD, and anti-β-actin antibodies. Cropped blots are displayed and full-length blots are presented in Supplementary Fig. S6. Bands corresponding to α-chain (b), β-chain (c), and ICD (d) of LRP1 and β-actin were scanned, after which the scanned bands were normalized by referring to the untreated control on the same blot. β-Actin was used as a loading control. Results are the means ± SD (n = 3 independent experiments). *Indicates a significant difference between 0 µM and 30 µM NMDA treatments without furin inhibitor (p < 0.05). (e) Effect of furin inhibitor on localization of α-chain (red) and β-chain of LRP1/ICD (green) at 4 h after 0 µM (control) or 30 µM NMDA (NMDA) treatment with or without 10 µM furin inhibitor. Representative images are shown from one experiment. The scale bar represents 30 µm. (f) The number of cells where LRP1-ICD was localized in the perinuclear region was counted. Results are expressed as the means ± SD of 4 independent experiments. *Significant difference from the NMDA- and furin inhibitor-untreated group (NMDA [0] and furin inhibitor [−]) (P < 0.05). ^#Significant difference from the NMDA-treated and furin inhibitor-untreated group (NMDA [30] and furin inhibitor [−]) (P < 0.05). The range of the number of counted cells in NMDA [0]-furin inhibitor [−], NMDA [0]-furin inhibitor [+], NMDA [30]-furin inhibitor [−], and NMDA [30]-furin inhibitor [+], were 14–36/experiment (the total number of cells counted: 110), 24–38/experiment (total: 128), 38–73/experiment (total: 216), and 27–57/experiment (total: 151), respectively. The total number of cells counted in Fig. 7f was 605.