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. 2019 Oct 7;10(10):761. doi: 10.1038/s41419-019-1992-4

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© The Author(s) 2019

Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/.

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Fig. 2 — Cells were treated with CCF or ICF in serum-free media for 24 h and cells cultured in the same condition without mechanical force application were used as the control (a). The mRNA expression of osteogenic marker genes was examined using real-time polymerase chain reaction (b–g). OSX protein expression was evaluated using immunofluorescence staining (h). In the inhibition experiments, cells were pretreated with inhibitor 30 min prior to ICF stimulation. Experimental conditions were illustrated (i). The OSX mRNA (j, n) were determined using real-time polymerase chain reaction. Osterix protein expression was evaluated using western blot (k, o) and the normalized band density was demonstrated (l, p). In addition, osterix protein expression was also determined by immunofluorescence staining (m, q). Bars indicate a significant difference between conditions. Scale bars indicate 50 μm

Fig. 2 — Cells were treated with CCF or ICF in serum-free media for 24 h and cells cultured in the same condition without mechanical force application were used as the control (a). The mRNA expression of osteogenic marker genes was examined using real-time polymerase chain reaction (b–g). OSX protein expression was evaluated using immunofluorescence staining (h). In the inhibition experiments, cells were pretreated with inhibitor 30 min prior to ICF stimulation. Experimental conditions were illustrated (i). The OSX mRNA (j, n) were determined using real-time polymerase chain reaction. Osterix protein expression was evaluated using western blot (k, o) and the normalized band density was demonstrated (l, p). In addition, osterix protein expression was also determined by immunofluorescence staining (m, q). Bars indicate a significant difference between conditions. Scale bars indicate 50 μm