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. 2020 Feb 18;26(6):1808–1831. doi: 10.1038/s41380-020-0671-2

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© The Author(s) 2020

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. 1 — a Timeline of drug treatment and behavioral testing. b Immunostaining of Saline + CTRL, MIA + CTRL, Saline + MG-REP and MIA + MG-REP animals at P42, after 3 weeks of PLX, and P60 after 18 days for repopulation, in the medial prefrontal cortex (mPFC), with antibodies against IBA1 (green) and DAPI (blue). Scale bar = 10 μm. c Quantification of IBA1⁺ cells in the mPFC showing complete microglial depletion by CSF1R inhibitor treatment and full recovery of microglia at P60. n = (3/2, 3/2, 4/2, 3/2) for male mice/litters per group P42 and n = (7/5, 7/5, 6/5, 6/3) male mice/litters per group for p60 (Saline + CTRL, MIA + CTRL, Saline + PLX and MIA + PLX). There was a main effect of prenatal treatment F(1,9) = 16.81, p = 0.0027 as well as main effect of drug treatment F(1,9) = 1486 p < 0.0001 and interaction effect F(1,9) = 16.31, p = 0.0029, at P42. d MIA male offspring show repetitive behavior deficits as shown in increased time spent grooming. Prenatal treatment effect, F(1,90) = 11.62, P = 0.0010, Drug effect, F(1,90) = 3.525, p = 0.0637, Interaction effect, F(1,90) = 0.5208, p = 0.4724, n = (23/9, 24/9, 19/11, 28/7) male mice/litters for (Saline + CTRL, MIA + CTRL, Saline + MG-REP, MIA + MG-REP). e–g Corrected social behavior in MIA male offspring by MG-REP: e Representative heatmaps of Three Chamber Social Interaction Test, “S”: tube containing social mouse, “E”: empty tube. f Difference in duration of time spent in “social” versus “opposite” chambers in saline offspring (left) and MIA offspring (right). Prenatal treatment effect, F(1,49) = 0.1947, P = 0.6610, Drug effect, F(1,49) = 5.289, p = 0.0258, chamber effect, F(1,49) = 57.20, p < 0.0001, Interaction effect, F(1,49) = 5.296, p = 0.0257. g Difference in duration of time spent sniffing the “social” versus empty “opposite” cylinders in saline offspring (left) and MIA offspring (right). Prenatal treatment effect, F(1,50) = 3.632, P = 0.0624, Drug effect, F(1,50) = 6.354, p = 0.0149, chamber effect, F(1,50) = 87.69, p < 0.0001, Interaction effect, F(1,50) = 1.544, p = 0.2198. n = (15/5, 15/9, 12/5, 15/4) male mice/litters for (Saline + CTRL, Saline + MG-REP, MIA + CTRL, MIA + MG-REP), *p < 0.05; **p < 0.01; ****p < 0.0001, ns denotes no significance; determined by 3-way ANOVA (alpha = 0.05) and Sidak post-hoc. Graphs indicate mean ± s.e.m.