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. 2020 Jul 20;30(10):885–901. doi: 10.1038/s41422-020-0370-1

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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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Fig. 4 — a, b qPCR analysis of Wnt7b, Wnt9a and Wnt3a in H2-c-fosLTR bones, tumor-bearing bones and dissected tumors (a) and primary OS cells isolated from Wls^WT-OS and Wls^ΔOB-OS mice (b). Bones from WT littermates (a) and MC3T3-E1 osteoblastic cells (b) were included for comparison. N.D., not detectable. c, d Gene expression in MC3T3-E1 cells expressing c-Fos-ER or mutant (inactive) c-Fos*-ER in the presence/absence of tamoxifen (c) and in the H2-c-fosLTR OS cell line (Fos^Tg-C3) expressing IPTG-inducible c-fos shRNA or non-target shRNA in the presence/absence of IPTG (d) was determined by qPCR. FosL1 (encoding Fra-1 and a bona fide Fos-target gene) is included as a control. e ChIP-qPCR quantification of c-Fos/AP-1 binding to Wnt7b and Wnt9a promoters in H2-c-fosLTR OS cells expressing IPTG-inducible c-fos shRNA or non-target shRNA in the presence/absence of IPTG. The FosL1 genomic region with a c-Fos/AP-1 binding site is included as a control. f Luciferase reporter assay in MC3T3-E1 cells cotransfected with increased amounts of c-Fos expressing vector (CMV-c-Fos) and Wnt7b and Wnt9a reporter constructs containing the ChIP sites from e. A Fosl1 reporter construct is included as a control. Bar graphs represent mean ± SEM. *P < 0.05 and **P < 0.01.