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. 2019 Aug 21;10:3758. doi: 10.1038/s41467-019-11674-z

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© The Author(s) 2019

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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Fig. 6 — Microglia facilitate plaque formation and compaction. a, b, Representative hemisphere stitches of 5xFAD and 5xFAD + PLX5622 mice stained for dense-core plaques (Thio-S in green) and immunolabeled for microglia (IBA1 in red). Scale bar = 1000 μm. c, d Confocal images of subicula stained with Thio-S for dense core plaques (green) and immunolabeled with IBA1 for microglia (red) and 6E10 for diffuse plaques (blue). Scale bar = 75 μm. e Quantification of IBA1⁺ cells in the subiculum (WT v 5xFAD, p < 0.001; WT v PLX5622, p < 0.001; PLX5622 v. 5xfAD + PLX5622, p = 0.002; 5xFAD v 5xFAD + PLX5622, p < 0.001). Two-way ANOVA with Tukey’s post hoc test; n = 7 for Wild-type, n = 6 for PLX5622, n = 6 for 5xFAD, n = 6 for 5xFAD + PLX5622. f, Plaque number within the subiculum is reduced by 33% in 5xFAD + PLX5622 mice compared to 5xFAD animals (p < 0.001). Two-tailed independent t-test; n = 6 for 5xFAD, n = 6 for 5xFAD + PLX5622. g–i Confocal images of dense-core plaques (Thio-S in green) and microglia (IBA1 in red) in the cortex, thalamus, and subiculum, respectively, of both 5xFAD groups showing an alteration in plaque morphology with CSF1R inhibitor treatment. Scale bar = 25 μm. Arrows point to zoomed images of dense-core plaques. Scale bar = 10 μm. j, k, Quantification of plaque circularity (j; 5xFAD v 5xFAD + PLX5622, p < 0.001) and Thio-S fluorescence intensity (k; 5xFAD v 5xFAD + PLX5622, p < 0.001). Two-tailed independent t-test; n = 6–7 for 5xFAD, n = 6 for 5xFAD + PLX5622. l–m Representative images from 7-month-old cohort immunolabled for ApoE (in red) and microglia (IBA1 in green) and stained for dense-core deposits (Thio-S in blue). Scale bar = 100 μm. n Quantification of ApoE immunoreactivity in plaque cores (cortex: p < 0.001; thalamus: p < 0.001). Two-tailed independent t-test; n = 7–10 for 5xFAD, n = 7 for 5xFAD + PLX5622. o–r, Immunolabeling and quantification of LAMP1 (o, p; p = 0.020) and APP (q, r; p = 0.068). Two-tailed independent t-test; n = 5 for 5xFAD, n = 4–5 for 5xFAD + PLX5622. Scale bar = 100 μm. Statistical significance is denoted by *p < 0.05, **p < 0.01, and ***p < 0.001. Statistical trends are denoted by ^#p < 0.10. Error bars indicate SEM