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. 2015 Oct 28;53(9):5912–5927. doi: 10.1007/s12035-015-9486-6

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© The Author(s) 2015

Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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.

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Fig. 7 — Conditioned medium from wild-type astrocytes expressing C/EBPδ promotes migration of the inactive astrocytes through MMP-3 activation. a In a wound healing assay, IL-1β-untreated, inactive wild-type astrocytes grow in conditioned media obtained from wild-type or C/EBPδ ^−/− astrocytes pretreated with or without IL-1β. Astrocytes show enhanced migration when cultured in the conditioned medium collected from IL-1β-pretreated wild-type astrocytes, suggesting a promoting effect of this conditioned medium on migratory behavior. b qPCR and Western blots reveal that the expression of both MMP-3 mRNA and protein increases significantly in wild-type astrocytes treated with IL-1β for 6 h. p84 is used as the loading control for Western blots. c A reporter assay shows that, when wild-type or C/EBPδ ^−/− astrocytes are transfected with the MMP-3 reporter, luciferase activity increases significantly in IL-1β-treated wild-type astrocytes. d A ChIP assay demonstrates that, in IL-1β-treated wild-type astrocytes, immunoprecipitated product (Q) captured by anti-C/EBPδ antibody contains both C/EBPδ and its binding target on the MMP-3 promoter, which is evidenced by the PCR, suggesting a direct binding of C/EBPδ to the MMP-3 promoter. e Transverse sections of the spinal cord immunostained with anti-GFAP and -MMP-3 antibodies demonstrate that wild-type mice exhibit higher MMP-3 immunoreactivity in the penumbral region (asterisks) lying between the lesion epicenter and the less injured tissue compared with the C/EBPδ ^−/− mice 28 days after SCI. f A scratch wound assay of wild-type astrocytes shows that the conditioned medium harvested from IL-1β-treated shLacZ-knockdown astrocytes significantly enhances astrocyte migration, which can be attenuated by the conditioned medium collected from IL-1β-treated shMMP-3-knockdown astrocytes (three left columns). Altered migratory behavior of these astrocytes is consistently associated with their expression of MMP-3 mRNA (three right columns)