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. 2020 May 19;10:8239. doi: 10.1038/s41598-020-65067-0

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© The Author(s) 2020

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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State-specific VDR protein expression of SMSCs in response to 1α,25(OH)₂D₃ treatment. (A) Representative immunoblot of VDR protein expression of developmental (D), mature (M), and aged (A) SMSCs after vehicle-treated and 100 nM 1α,25(OH)₂D₃-treated for 48 h (daily-treated). The highly sensitive VDR (D-6) antibody was used to detect VDR protein expression and GAPDH served as loading control. K = Kidney and I = Intestine (positive controls). VDR protein expression level was normalized with GAPDH protein expression that obtained from the same gel and experiment. Data obtained from 1α,25(OH)₂D₃-treated group was normalized with the respective vehicle-treated group to demonstrate the fold change at each growth stage (n = 3 SMSCs isolated from different mice/growth stage). *p < 0.05 compared to vehicle-treated group (developmental SMSC). (B) VDR protein expression (arrows) in developmental SMSC (MyoD⁺ cells) was substantial increased after treated with 100 nM 1α,25(OH)₂D₃ for 48 h (daily-treated). (C) VDR protein specifically expressed in undifferentiated SMSC (Myogenin^- cells) (arrows) after treated with 100 nM 1α,25(OH)₂D₃ for 48 h (daily-treated). DAPI was used to visualize nuclear localisation. Image (B,C) were taken at ×200 and ×400 magnifications. Rectangular line in (B) represents the area that is illustrated at ×400 magnification.