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. 2019 Jun 13;10:2588. doi: 10.1038/s41467-019-10411-w

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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. 7 — Phagocytosis of apoptotic cells by activated microglial in Knl1 cKO and Knl1/Trp53 dKO. a Developmental analysis by DAPI staining revealed that pyknotic nuclei (open arrowheads) were present at E13.5, peaked in number at E15.5, and eliminated by birth in cKO cortex (mean, two-tailed unpaired t-test, E13.5, ctrl: n = 4 cKO: n = 3, E15.5, ctrl: n = 4, cKO: n = 6, E17.5, n = 3, P0, ctrl: n = 4, cKO: n = 3, P4, n = 4 animals, scale bar: 50 µm). b ADGRE1 (F4/80) immunostaining revealed morphologic activation of microglia in E15.5 cKO and dKO cortex (scale bar: 100 µm). Confocal z-stacking revealed that ADGRE1+ (green) microglia acquired a ballooned morphology. c ADGRE1+ (green) microglia with ballooned morphology phagocytosed dozens of DAPI+ (white) pyknotic nuclei, some of which were CC3+ (red, and open arrowheads) in cKO and dKO cortex at E15.5 (scale bar: 5 µm). d Phagocytic microglia in cKO and dKO were significantly larger in size compared to ctrl microglia. However, the increase in microglial size was significantly attenuated in dKO compared to cKO (mean, ANOVA with post-hoc t-test, n = 3 animals). e The number of AIF1+ (IBA1+) microglia was significantly increased in cKO and dKO compared to ctrl. The increase in microglial number, however, was attenuated in dKO compared to cKO (mean, ANOVA with post-hoc t test, ctrl: n = 5, cKO: n = 4, dKO: n = 3 animals)