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. 2021 Oct 11;22(20):10954. doi: 10.3390/ijms222010954

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© 2021 by the authors.

Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).

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Immunostaining of AR S210/213 and AKT S473 in U87MG subclones. (A) U87MG WT, U87MG EGFR, and U87MG EGFRvIII cells were treated with vehicle (1% DMSO), 5 μM afatinib, or 5 μM MK2206 for 6 h and analyzed by immunofluorescence with antibodies against pAR (S210/213) (red) and pAKT (S473) (green). (B,C) AR nuclear localization was determined by quantifying the relative fluorescent intensity ratio between nuclear and cytosolic compartments using ImageJ software. The results are expressed as mean ± SD following normalization to U87MG WT (B) or the vehicle-treated subclones (C); p-values between the groups are indicated.