Supplementary Data from Targeting Tissue Factor for Immunotherapy of Triple-Negative Breast Cancer Using a Second-Generation ICON

S1. Tissue factor expression on TNBC and non-TNBC lines with BRCA1/2-wild type or mutation by flow cytometry assay. S2. TF expression is detected on the TNBC cancer cells and tumor vascular endothelial cells, but not on adjacent normal tissues, on TNBC CDX and PDX tumors. S3. Immunohistochemical (IHC) staining for TF and CD31 expression on paraffin-embedded whole TNBC tissues and tissue-microarray slides (TMA). S4. Schematic structures of tissue factor-targeting second-generation ICONs (murine and human L-ICON1s) and their binding activities to human and murine breast cancer cells. S5. Coagulation activities of L-ICON1 and ICON determined by Factor VII Human Chromogenic Activity Assay. S6. Generation of a murine breast cancer line 4T1 stably expressing luciferase (Luc) and green fluorescent protein (GFP) and an orthotopic mouse model of murine TNBC. Table S1. Coagulation activities (IU/ml, mean {plus minus} SD) of L-ICON1, ICON(WT) and ICON(K341A). S2. Protein concentrations of L-ICON1 and ICON in vitro in 4-day cell culture serum free medium of adenoviral vector-infected human TNBC MDA-MB231 line and in vivo in mouse plasma two days after last intratumoral injection of Ad vectors in an orthotopic SCID mouse model of human MDA-MB-231 tumor xenograft.