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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) Effect of furin inhibitor on localization of TGN46 and LRP1-ICD at 4 h after 0 µM (control) or 30 µM NMDA (NMDA) treatment of cortical neurons with or without 10 µM furin inhibitor. The cells were immunostained with anti-TGN46 and anti-β-chain/ICD antibodies. Representative images are shown from one experiment. The scale bar represents 30 µm. (b) The number of LRP1-ICD immune-positive cells that co-localized with TGN46 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 29–40/experiment (the total number of cells counted: 133), 22–27/experiment (total: 98), 51–82/experiment (total: 186), and 20–32/experiment (total: 109), respectively. The total number of cells counted in Fig. 8b was 526. (c) Effect of furin inhibitor on localization of furin and LRP1-ICD at 4 h after 0 µM (control) or 30 µM NMDA (NMDA) treatment with or without 10 µM furin inhibitor. The cells were immunostained with anti-furin and anti-β-chain of LRP1/ICD antibodies. Representative images are shown from one experiment. The scale bar represents 30 µm. (d) The number of LRP1-ICD immune-positive cells that co-localized with furin in the perinuclear region was counted. Results are expressed as the means ± SD of 4 independent experiments. *Significant difference from the NMDA- and furin-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 17–27/experiment (the total number of cells counted: 91), 17–33/experiment (total: 110), 27–63/experiment (total: 179), and 19–40/experiment (total: 111), respectively. The total number of cells counted in Fig. 8d was 491.