JNCI Journal of the National Cancer Institute

The following supplementary figure accompanies this article

Files in this Data Supplement:

• Supplementary Figure 1 - Supplementary Fig. 1 - Endorepellin preparation purity, lack of tumor localization when inactivated, and effects on tumor VEGF expression. A) Approximately 5 μg of purified endorepellin (ER) was run on 8% SDS-PAGE and bands were resolved by staining with colloidal Coomassie blue (Invitrogen) for 7 hours, and destaining in water for 18 hours. Note that no peptide bands beyond endorepellin and LG3 (a proteolytic fragment of endorepellin) are present. B) Coomassie stained 8% SDS-PAGE gel showing native and heat inactivated (H.I.) endorepellin protein. Note that both proteins migrate similarly in the gel other than a small band located above 250 kDa, representing a small percentage of the heat inactivated endorepellin that forms high molecular weight aggregates upon cooling. C) Tumor endothelial cells (TEC) isolated from A431 tumors labeled with the endothelial cell-specific marker Bandeirea Simplicifolia Lectin I-B4 (conjugated with FITC) D) Representative DAPI counterstained sections of A431 tumors after chronic treatment with either heat inactivated (H.I.) or native endorepellin immunostained with anti-his and anti-PECAM (CD31) demonstrating the lack of H.I. endorepellin deposition in the tumor vasculature, in contrast to the native endorepellin which targets the tumor vasculature. In these experiments, treatments began 24 hours after tumors became visible. E) Representative section of A431 tumor xenografts following chronic treatment with native endorepellin (with and without DAPI counterstain) immunostained with anti-VEGF and anti-CD31. Note the vesicular, perivascular VEGF signal in CD31 negative cells. Scale bar: 50 μm.