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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. 3 — Extended elimination of microglia does not induce peripheral leukocyte or behavioral abnormalities. 1.5-month-old wild-type (WT) or 5xFAD mice were treated with control chow or PLX5622 for 10 or 24 weeks. a Experimental design. b Terminal PK of PLX5622 and 5xFAD + PLX5622 groups. Two-tailed independent t-test. n = 5–6 for PLX5622, n = 8–11 for 5xFAD + PLX5622. c Hemisphere stitches of microglia (IBA1 in green) in the 7-month-old cohort. Scale bar = 1000 μm. d Analysis of different subsets of leukocytes (CD11b, p = 0.059; CD19, p = 0.052; all others NS). Two-tailed independent t-test; n = 4–5 for Wild-type, n = 3–4 for PLX5622. e–f All analyses listed in respective order for retrosplenial (RS) cortex, somatosensory (SS) cortex, and thalamus. Microglial number in 4 e and 7 f month-old cohorts (4-month cohort: WT v 5xFAD, p = 0.001, NS, p < 0.001; WT v PLX5622, p < 0.001, p < 0.001, p = 0.044; 5xFAD v 5xFAD + PLX5622, p < 0.001, p < 0.001, p < 0.001. 7-month cohort: WT v 5xFAD, p = 0.015, p = 0.024, p < 0.001; WT v PLX5622, p < 0.001 for all regions; 5xFAD v 5xFAD + PLX5622, p < 0.001 for all regions). Two-way ANOVA with Tukey’s post hoc test; n = 4–6 for Wild-type, n = 4–6 for PLX5622, n = 5–8 for 5xFAD, n = 6–9 for 5xFAD + PLX5622. g–h Measurements for anxiety were performed by Elevated Plus Maze (EPM) and quantified as number of arm entries (g WT v 5xFAD, NS (open) and p = 0.050 (closed); PLX5622 v 5xFAD + PLX5622, p = 0.007 (open) and p < 0.001 (closed), 5xFAD v 5xFAD + PL5622, NS (open) and p = 0.056 (closed)) and time in arms (h WT v 5xFAD, p = 0.0157 (open) and p = 0.046 (closed); PLX5622 v 5xFAD + PLX5622, p < 0.001 (open) and p < 0.001 (closed); 5xFAD v 5xFAD + PLX5622, p < 0.001 (open), p < 0.020 (closed)). i Mean latencies to a hidden platform from the Morris water maze (MWM) acquisition trials. j–k Number of platform entries (j; PLX5622 v 5xFAD + PLX5622, p = 0.071) and time in platform zone (k; WT v PLX5622, p = 0.094; PLX5622 v 5xFAD + PLX5622, p = 0.073) in the MWM probe trial. For all behavioral analyses: Two-way ANOVA with Tukey’s post hoc test; n = 8–12 for Wild-type, n = 8–12 for PLX5622, n = 9–12 for 5xFAD, n = 8–9 for 5xFAD + PLX5622. Statistical significance is denoted by *p < 0.05, **p < 0.01, and ***p < 0.001. Statistical trends are denoted by ^#p < 0.10. NS, not significant (p > 0.10). Error bars indicate SEM