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. Author manuscript; available in PMC: 2017 Sep 1.
Published in final edited form as: Matrix Biol. 2016 Mar 22;55:22–34. doi: 10.1016/j.matbio.2016.03.006

Figure 5.

Figure 5

Therapy-generated soluble heparanase activates pro-tumorigenic gene expression and cell signaling. (A) ELISA measuring the level of TNF-α secreted by J774 A.1 cells after incubation with Con CM or BTZ CM. Controls included BTZ CM immunodepleted of HPSE using HPSE IgG (HPSE IP) or Con IgG (Con IP). *p<0.001 versus Con CM, # p<0.01 versus Con IP. (B) Real-time PCR for VEGF, MMP-9, and HGF in RPMI-8226 cells after incubation with Con CM, BTZ CM immunodepleted with anti-heparanase antibody (HPSE IP) or BTZ CM immunodepleted with isotype matching control antibody (Con IP). Controls included Roneparstat (6.75uM) added along with BTZ CM (BTZ CM + Roneparstat). $p<0.05 versus Con CM, *p<0.05 versus BTZ CM + HPSE IP, # p<0.05 versus BTZ CM + Con IP. (C) For testing effect of sHPSE on cell signaling, we used concentrated conditioned media from BTZ treated CAG cells (BTZ CAG CM) and concentrated conditioned media from vehicle treated CAG cells (Con CAG CM). Western blot for different signaling molecules from myeloma cell lines after 2 h incubation; 1 - Con CAG CM, 2 - BTZ CAG CM, and 3 - BTZ CAG CM + Roneparstat (Rone, 6.75 μM).

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