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. 2019 May 9;9:7146. doi: 10.1038/s41598-019-43709-2

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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) p46-JNK and p54-JNK level measurement. p46-JNK was not affected by treatments while p54-JNK was increased after 10 min NMDA (100 μM) + glycine (1 μM) alone or pre-treated with JGRi1 (1 μM) or sJGRi1. JGRi1 (1 μM) or sJGRi1 (1 μM) alone did not affect JNK phosphorylation state. Means ± s.e.m. n = 7, ***p < 0.001 NMDA stimulus vs. control, **p < 0.01 NMDA + sJGRi1 vs. control; Newman-Keuls’s test. Cropped WBs are here shown. (B) NMDA stimulus increased STX1a phosphorylation that was instead prevented by pretreatment with JGRi1 (1 μM), but not by sJGRi1 (1 μM). Means ± s.e.m. n = 7, *p < 0.01 NMDA stimulus vs. control, ***p < 0.001 JGRi1 alone vs NMDA, ***p < 0.001 NMDA + JGRi1 vs NMDA, *p < 0.05 NMDA + sJGRi1 alone vs. control; Newman-Keuls’s test. Cropped WBs are here shown. (C) Where indicated JGRi1 (1 μM) was applied to cortical synaptosomes 30 min before 10 min NMDA (100 μM) + glycine (1 μM) stimulus. SNARE complex formation was assessed by probing STX1a in unboiled samples comparing STX1a lanes smearing (50 to 100 kDa) with free STX1a in a cropped WB. NMDA increased SNARE complex formation while JGRi1 prevented it. Means ± s.e.m. n = 4, ***p < 0.001 NMDA stimulus vs. control, **p < 0.01 JGRi1 alone vs NMDA; Newman-Keuls’s test.