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. Author manuscript; available in PMC: 2011 Nov 1.
Published in final edited form as: Cancer Res. 2010 Oct 26;70(21):8329–8338. doi: 10.1158/0008-5472.CAN-10-2179

Figure 5. Recombinant heparanase (rHPSE) induces RANKL expression and secretion in myeloma cells.

Figure 5

Equal numbers of human myeloma U266 or MM.1S cells were grown in complete medium in the absence or presence of rHPSE (100 ng/ml, added every 12 hours) for 48 h. The cells were lysed for western blotting, and the conditioned media were harvested for ELISA. A. Western blots were probed with antibody to human RANKL or β-actin. Saos-2 cell lysate was used as a positive control for human RANKL. B. RANKL bands from western blots were quantified using ImageJ and normalized to the corresponding β-actin bands. Each bar graph represents an average quantitative result of two independent experiments with two western blots. C. RANKL ELISA of conditioned media. The graph shows the percentage increase in RANKL present in medium conditioned by cells treated with rHPSE compared to control treated cells. The results are means ± SD from 2 independent experiments, *** p<0.001 to the controls.