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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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Western blot analysis of AR S210/213 and AKT S473 in U87MG subclones. (A) Cells were treated for 30 min (30′) to 6 h with 5 μM afatinib and analyzed with Western blotting using antibodies against AKT, pAKT (S473), and pAR (S210/213). The anti-GAPDH housekeeping protein was used as a loading control. (B) Protein fold change (Y-axis) of each sample compared to treatment at time zero was calculated according to band densitometry analysis with ImageJ software, following normalization to GAPDH.