Skip to main content
. 2016 Dec 27;8(12):19403–19412. doi: 10.18632/oncotarget.14292

Figure 2. Recombinant heparanase enzyme enhances insulin receptor signaling pathway in breast cancer cells.

Figure 2

A–C. Human breast carcinoma MCF-7 cells were serum-starved overnight and then either remained untreated (c) or stimulated with insulin (100 nM) for 30 min in the absence or presence of 0.8 μg/ml active recombinant heparanase (Hpa) or heat-inactivated heparanase (iHpa). A. Cell lysates containing equivalent amounts of total protein were then immunoblotted using antibody specific for phospho-insulin receptor (pINSR), phospho-AKT (pAKT), total INSR, total AKT or total actin. B, C. The band intensity was quantified using ImageJ software; intensity ratio for pINSR/total INSR (B) and pAKT/total AKT (C) are shown. The data are representative of three independent experiments. Inset: Enzymatic activity in samples of Hpa (black line) and iHpa (red line) was examined as described in Methods. D–F. Mouse E0771 breast cancer cells (characterized by low endogenous levels of heparanase) were serum-starved overnight and then treated as described in A-C. Intensity ratio for pINSR/total INSR (E) and pAKT/total AKT (F) are shown. The data are representative of three independent experiments.