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. 2018 Jun 29;8:9821. doi: 10.1038/s41598-018-28183-6

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© The Author(s) 2018

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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Mild C3 activation. (A) To evaluate an activation of C3, retinal cross-sections were stained with anti-C3 (red) and DAPI (cell nuclei, blue) 3, 7, and 14 after immunization. (B) No alterations in C3⁺ cell numbers were noted in the S100 group after 3 and 7 days (p > 0.05). After 14 days, significantly more C3 depositions were observed (p = 0.04). (C) The C3 mRNA expression level showed no alterations 3 and 7 days after immunization (p > 0.05). A significant higher level of C3 mRNA was observed at 14 days (p < 0.001). (D) Anti-C3 staining (red) of the optic nerves was performed 3, 7, and 14 days after immunization. Cell nuclei were labeled with DAPI (blue). (E) No changes were revealed at all points in time (p > 0.05). Abbreviation: GCL = ganglion cell layer. Values are mean ± SEM for immunohistology and median ± quartile + maximum/minimum for qRT-PCR. Scale bars: 20 µm.