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. 2022 Apr 12;19:91. doi: 10.1186/s12974-022-02456-4

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© The Author(s) 2022

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 2 — Pharmacological suppression of SPAK–NKCC1 complex phosphorylation in choroid plexus after ischemic stroke. a Experimental protocol. Mice were subjected to tMCAO and randomly assigned to vehicle (Veh), ZT-1a or bumetanide (BMT) treatment group administered via intraperitoneal injection (i.p.) with a half-dose at 3 h and 8 h post-reperfusion (Rp), respectively. b Representative infarct images with MAP2 expression staining at 24 Rp. c Infarct volume quantification summary. Data are mean ± SD (n = 6, 4 male, 2 female). *p < 0.05. One-way ANOVA. d Representative immunofluorescent images of pSPAK and pNKCC1 staining in the Sham or ipsilateral (IL) LVCP post-stroke brains. Arrowheads: low level expression of pSPAK or pNKCC1. Arrows: elevated pSPAK or pNKCC1 expression. e Quantification summary. Data are mean ± SD (n = 6, 4 male, 2 female). *p < 0.05, ***p < 0.001, ****p < 0.0001. One-way ANOVA. f Western blot analysis of SPAK–NKCC1 cascade expression in the ChP of Sham, stroke Veh-control, ZT-1a- or BMT-treated mice. ChP tissue lysates were subjected to immunoblotting with the indicated antibodies. g Immunoblot summary. Data are mean ± SD (n = 4, 2 male, 2 female). *p < 0.05, **p < 0.01, ***p < 0.001. One-way ANOVA