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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) Schematic drawing of brain regions used in the experiments. (b) Changes in α-chain, β-chain, and ICD of LRP1 at 24 h after MCAO/R. Proteins from non-ischemic (N) and ischemic (I) areas from the ipsilateral (ipsi) and contralateral (cont) cortex in MCAO/R rats were analyzed by Western blotting with anti-α-chain and anti-β-chain/ICD of LRP1, and anti-β-actin antibodies. Cropped blots are displayed and full-length blots are presented in Supplementary Fig. S1. Bands corresponding to α-chain (c), β-chain (d), and ICD (e) of LRP1 and β-actin were scanned, and the scanned bands were normalized by the untreated naïve control on the same blot. β-Actin was used as a loading control. Results are the means ± SD (n = 3 rats per group). *Indicates a significant difference from the corresponding area of the contralateral hemisphere (p < 0.05). (f) Changes in localization of β-chain and/or ICD of LRP1 and TGN at 24 h after MCAO/R. Non-ischemic and ischemic areas of the ipsilateral and contralateral hemispheres in the brain of MCAO/R rats were immunostained with anti-TGN46 and anti-β-chain/ICD of LRP1 antibodies, and then observed with a confocal microscope. The colocalization of TGN46 with LRP1-ICD is indicated by the “white arrow”. Representative images are shown from one rat. The scale bar represents 30 µm. (g) Changes in localization of β-chain and/or ICD of LRP1 and TGN at 24 h after MCAO/R. The number of LRP1-ICD immune-positive cells that co-localized with TGN46 was counted. Results are expressed as the means ± SD (n = 4 rats per group). *Indicates a significant difference from the corresponding area of the contralateral hemisphere (P < 0.05). The range of the number of cells counted for N-cont, I-cont, N-ipsi, and I-ipsi were 22–60/rat (the total number of cells counted: 150), 14–66/rat (total: 139), 22–59/rat (total: 159), and 23–63/rat (total: 156), respectively. The total number of cells counted in Fig. 1g was 604.