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. 2016 Nov 16;6:37184. doi: 10.1038/srep37184

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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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(A) Small hairpin RNAs (shRNA) were used to knock down Cx43 expression in MC-3T3 cells, and a scrambled sequence was used as a control. The expression of Cx43 was detected by Western blot. (B) MC-3T3 cells transfected with Cx43 shRNA were induced in osteogenic medium. Alizarin Red S (week 2) and ALP (week 1) staining was performed. (C) Calcium deposits and ALP activity were quantified. (D) mRNA expression of osteogenic marker genes was detected by real-time PCR at week 1 and 2. (E) ERK pathway activation and Cx43 expression were examined weekly by Western blot. (F) A densitometric analysis of p-ERK expression was performed (t-test, n = 3, *p < 0.05; NS, not significant).