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. 2019 Feb 18;19:158. doi: 10.1186/s12885-019-5363-4

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© The Author(s). 2019

Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided 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 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.

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Fig. 1 — (a) gnrh (b) kiss1, (c) gnrhr, and (d) gpr54/kiss1r mRNA expression in normal canine osteoblasts (OPC), multiple osteosarcoma cell lines (COS, POS, HMPOS, C4, D17, human SAOS) and clinical patient-derived samples (1014, 1036). Arrow in (b) indicates kiss1 band. PCR primers used can be found in Additional file 2 Table S1. e β-actin (actb) expression in OPC, COS, POS, HMPOS, and D17 cells. f Relative levels of cKiss1R expression in COS vs. OPC. g Immunoblot of Kiss1R protein (54kD, upper band) in normal canine osteoblasts (OPC) and osteosarcoma lines (COS, POS); α-tubulin (42kD, lower band) was also probed as a control. h RT-PCR expression of rank in COS vs. OPC. i RT-PCR expression of rankl in COS vs. OPC (3 preparations). NTC indicates RNA (no cDNA template) control