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. 2018 Feb 1;8:2134. doi: 10.1038/s41598-018-20571-2

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© The Author(s) 2018

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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GPR54 is necessary for KP-10-induced osteoblast differentiation. (A) Wild-type and GPR54^−/− C3H10T1/2 cells were treated with 50 μM KP-10 for 6, 9, 12 or 24 h. Western blot analysis was performed using the indicated antibodies. BMP2 expression was normalized to β-actin expression. And the densitometry analysis was performed by ImageJ program (A, lower panel). (B and C) RT-PCR was performed using total RNA isolated from cultured cells. Wild-type and GPR54^−/− cells were treated with 50 μM KP-10 for 1 day (B and C). C.M of wild-type cells treated with 50 μM KP-10 for 12 h was collected. GPR54^−/− C3H10T1/2 cells were treated with 50 μM KP-10 or C.M for 2 days (C).