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. 2018 May 3;9(5):512. doi: 10.1038/s41419-018-0545-6

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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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Fig. 4 — a–d The expressions of Dicer1 mRNA and protein were dramatically decreased compared to that of the control group. And the protein expression of p21 increased. e Representative micrographs after SA-β-gal staining of Dicer1-KD MSC (shRNA), negative MSC (transfected with control lentiviruses) and control-MSC (HC-MSC without transfection) (100× magnification). f One hundred MSC per sample were counted using light microscopy, and the percentages of SA-β-gal-positive cells were determined. The average of three replicates is displayed. g The proliferation of MSCs treated with Dicer1 knockdown (KD) was obviously inhibited in comparison with either control MSCs or the negative group. h Cell cycle analysis of Dicer1-KD MSC by flow cytometric analysis. Dicer1 KD caused an increasing proportion of cells in the G1 phase and a decrease of those in the S phase without inducing apoptosis. i After 21 days of osteogenic induction, Alizarin red S staining was performed to visualize osteogenic differentiation. Representative original images of BMMSCs derived from control-MSC (HC-MSC without transfection), negative MSC (transfected with control lentiviruses), Dicer1-KD MSC are shown. j Relative calcium production (OD 572 nm) by Dicer1-KD MSC, was significantly lower after 21 days of differentiation as compared with controls. k The ALP activity of Dicer1-KD MSC was significantly lower than that of controls after 3 days culturing in osteogenic medium (OM). l, m Relative RUNX2 and ALP mRNA expression levels. The average of three replicates is displayed. Compared with controls, the significance was set as * p ≤ 0.05; ** p ≤ 0.01