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. 2016 Sep 7;6:32771. doi: 10.1038/srep32771

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Copyright © 2016, The Author(s)

This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/

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AR and osteoactivin coding sequences (CDS) were expressed in α1-AMPK MC3T3-E1 (A), primary osteoblasts (G) and mouse BMSCs (L) and their expression was knocked down in α2-AMPK MC3T3-E1 (A), primary osteoblasts (G) and mouse BMSCs (L). An empty LV vector (LV-ctr) and a vector expressing scrambled siRNA (LV-scr) served as the respective controls. MC3T3-E1, primary osteoblasts and mouse BMSCs were induced to osteogenic differentiation. On day 7 (MC3T3-E1 and mouse BMSCs) and day 9 (primary osteoblasts) after induction, the expression of the osteogenesis markers Runx2 (B,M), Osterix (H), Alp (C,I,N), Ibsp (D,J,O), and Ocn (E,K,P) was evaluated by qRT-PCR. β-actin was used as an internal control of qRT-PCR. On day 21 after induction, calcium deposition was assessed by alizarin red staining and quantification (F). Data are shown as mean ± SD; *P < 0.05.