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. 2023 Nov 18;40(4):483–499. doi: 10.1007/s12264-023-01147-1

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© The Author(s) 2023

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 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/.

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Fig. 7 — MFG-E8 is mainly expressed and secreted by astrocytes in the central neuronal system. A Representative immunofluorescence co-staining images of MFG-E8/Iba-1, MFG-E8/GFAP, MFG-E8/Oligo2, and MFG-E8/Map2 in the CC of WT sham mice. Microglia, astrocytes, oligodendrocytes, and neurons are labeled with Iba-1, GFAP, Oligo2, and Map2, respectively. White scale bar: 50 μm; orange scale bar: 10 μm. B Gating strategy for flow cytometry and FACS-sorting. C The mRNA levels of MFG-E8 in the microglia, astrocytes, and other cells from WT mice in the Sham and BCAS groups by qPCR (n = 4 per group; ***P < 0.001 vs astrocytes in sham group, ^##P < 0.01 vs astrocytes in BCAS group, two-way ANOVA). D MFG-E8 mRNA expression in primary neurons, oligodendrocytes, microglia, and astrocytes without any treatment (n = 4 repeats per group; *P < 0.05, ****P < 0.0001 vs neurons, ANOVA). E, F Immunoblot bands (E) and quantification (F) of MFG-E8 expression in primary microglia and astrocytes with GAPDH as a loading control (n = 3 repeats per group; ***P < 0.001 vs microglia, unpaired t-test). G MFG-E8 mRNA expression in primary astrocytes at different time points after myelin stimulation (n = 3 per repeats group; *P < 0.05 vs PBS group, ANOVA). H, I Representative immunoblots (H), and densitometry analyses (I) of MFG-E8 expression in primary astrocytes at different time points after myelin stimulation (n = 3 per repeats per group; *P < 0.05, **P < 0.01 vs PBS group, ANOVA). J, K Representative immunoblots (J), and densitometry analyses (K) of MFG-E8 expression in primary astrocytes at different time points after CoCl2 stimulation (n = 3 repeats per group; **P <0.01, ***P < 0.001, ****P < 0.0001 vs PBS group, ANOVA. All data are presented as the mean ± SD).