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. 2020 Apr 30;10:7354. doi: 10.1038/s41598-020-64363-z

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© The Author(s) 2020

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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CNO-gated DREADD treatment suppressed EAE severity. (a) Schematic illustration of pAAV-CaMKIIα-hM4Di (Gi)-mCherry (Gi-DREADD) and pAAV-CaMKIIα-EGFP (control-DREADD). The AAV9 vector contains Gi- or control-DREADD under the CaMKIIα promoter. (b) Distribution of control- and Gi-DREADD at spinal level Th 8 in mice 14 days after injection of DREADD-carrying-AAV9. Scale bars: 150 μm. (c) Higher magnification images showing DREADD colocalization with NeuN or GFAP in the dorsal column of the Th 8 level spinal cord. NeuN (red) and GFAP (magenta) in control-DREADD (green) experiment. NeuN (green) and GFAP (cyan) in Gi-DREADD (red) experiment. Scale bar: 50 μm. (d) Experimental schema. Targeted EAE was induced with MOG_35–55 peptide and local injection of TNFα and IFNγ. Seven days after MOG immunization, DREADD-carrying-AAV9 was injected at spinal level Th 9. EAE scores were assessed from day 0 to 28 after cytokine injection. (e) EAE scores throughout the 28 days (control: n = 8, Gi: n = 9; two-way ANOVA followed by the Sidak test). (f) Maximum and (g) accumulated EAE scores (control: n = 8, Gi: n = 9; Student’s t-test). Data are presented as mean ± sem.