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. 2015 Aug 11;11(9):1594–1607. doi: 10.1080/15548627.2015.1068489

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© 2015 The Author(s). Published with license by Taylor & Francis Group, LLC

© Phillip T Newton, Karuna K Vuppalapati, Thibault Bouderlique, and Andrei S Chagin

This is an Open Access article distributed under the terms of the Creative Commons Attribution-Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/), which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. The moral rights of the named author(s) have been asserted.

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Figure 6. — The stimulation of RPS6 phosphorylation by Baf is nutrition independent. C5.18 cells were differentiated into chondrocytes and then treated with Baf in the presence of different serum concentrations for 48 (A, B) and 24 (C, D) h. Elevation of RPS6 phosphorylation was observed independently of serum levels. (E, F) Amino acid- and glucose-free custom-made DMEM medium was supplemented with glucose (100% of standard DMEM) and nonessential amino acids (100% of standard DMEM), whereas the levels of essential amino acids (EAA) varied from 100% to 0% (of standard DMEM). Differentiated C5.18 chondrocytes were treated with Baf in serum-free conditions with varying levels of essential amino acids. Levels of SQSTM1, phospho-RPS6, total RPS6, phospho-EIF4EBP1 and total EIF4EBP1 were analyzed by western blot (*p < 0.05, n = 2–4).