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. 2018 Apr 26;6:14. doi: 10.1038/s41413-018-0015-x

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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 — Wnt7b and Bmp2 expression is increased in Lsd1 deleted osteoblasts. a–c Lsd1^fl/fl calvarial cells derived from P3 mice were infected with Cre-lentivirus or Egfp-lentivirus followed by culturing in osteoblast differentiation media for 14 days. ALP staining (day 7), alizarin red staining (day 14) (a) and quantification of ALP activity (b) of calvarial cells in osteogenic cultures. Data in b are the mean ± SD (n = 6), ***P < 0.001. Unpaired t-test was performed. c mRNA expression of osteoblast marker genes at day7 was analyzed by RT-PCR, n = 3, unpaired t-test, *P < 0.05, **P < 0.01. d Histone H3 lysine4, lysine9 methylation, and LSD1 levels were measured by western blot. e Venn diagram shows the overlap of genes of increased expression level and increased H3K4me2 occupancy at the TSS region in LSD1 mutant osteoblasts. f Gene Ontology (GO) analysis within the 114 genes using the database for annotation, visualization, and integrated discovery (DAVID) bioinformatics tools. The enriched GO biological processes were identified and listed. The enrichment P-value is <0.01. g RT-PCR analysis of the expression levels of candidate genes in RNA-seq samples. Data in g represent the mean ± SD (n = 3) Unpaired t-test was performed. *P < 0.05, **P < 0.01, ***P < 0.001.