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. 2019 Jul 25;10:1738. doi: 10.3389/fimmu.2019.01738

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Copyright © 2019 DuBois, Ray, Gruber, Zhang, Aflakpui, Macian-Juan and Shafit-Zagardo.

This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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Conditional knockout of Akt3 in CD4⁺ T-cells results in earlier disease onset during MOG-induced EAE. (A) Western blot analysis of protein homogenates of lymph node CD4⁺ T-cells, brain, and spinal cord isolated from (1) Akt3^−/−, (2) CD4-CKO, and (3) Akt3^fl/fl. Densitometry analysis of CD4-CKO and Akt3^fl/fl brain and spinal cord homogenates yielded approximately equal ratios of Akt3/β-actin. (B) EAE clinical course and (C) day of disease onset (CI ≥ 1) of CD4-CKO mice (n = 7) and Akt3^fl/fl (n = 9), representative graph of 4 independent experiments. (D) Histological analysis of microglia/macrophages (Iba1)—quantified in (E), T-cell infiltrates (CD3)—quantified in (F), axonal damage (SMI32) quantified in (G), and (H) demyelination (MBP) in lumbar spinal cord of CD4-CKO and Akt3^fl/fl controls after 5 consecutive days with clinical scores (*p < 0.05, Mann-Whitney U-test). Scale bars = 200 μm.