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. 2021 Jan 4;22:1. doi: 10.1186/s13059-020-02207-9

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© The Author(s) 2021

Open AccessThis 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 licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence 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 licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.

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Fig. 4 — Functional assays in human mesenchymal stem cell-induced osteoblasts following siRNA knockdown of four genes each at three loci implicated by GWAS and variant-to-gene mapping. a, e, i Representative alkaline phosphatase (blue) and Alizarin Red S (red) stains for osteoblastic activity and calcium deposition, respectively, for the three tested loci. Experiments were performed twice in three unique donor lines. b, f, j Quantification of alkaline phosphatase staining using quantitative image analysis was repeated twice with three different independent hMSC donor cell lines. c, g, k ALPL gene expression. d, h, l Quantification of Alizarin Red S staining. *p < 0.05 comparing no treatment to BMP treatment for each siRNA, ^#p < 0.05 comparing control siRNA to siRNA for gene of interest, n.s = not significant. Error bars represent standard deviation. hMSC donor line, siRNA, and qPCR details are given in Additional file 1: Tables S19-S21