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