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. 2012 Aug;27(6):384–391. doi: 10.1089/cbr.2012.1209

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Copyright 2012, Mary Ann Liebert, Inc.

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FIG. 2. — Binding and internalization of hlFVII-LDP. (A) The binding specificity of hlFVII-LDP to TF expressed on HepG2 cells was verified by coimmunoprecipitation assay. Total protein extracted from HepG2 cells (lanes 1, 3, and 4) and MCF-7 cells (lane 2) incubated with bovine serum albumin (lane 1) or hlFVII-LDP (lanes 2–4) was incubated with either preimmune serum (lane 4) or an antihuman TF monoclonal antibody (lanes 1–3). The complexes formed were collected with Protein A agarose and were analyzed by SDS-PAGE and Western blot with antihuman FVII monoclonal antibody. (B) Expression of human TF on different cancer cell lines analyzed by Western blot with antihuman TF monoclonal antibody. (C) Internalization assay of hlFVII-LDP by hlFVII-LDP-mCherry. Cells in 1–3 and 7–9 were treated with mCherry, and cells in 4–6 and 10–12 were treated with hlFVII-LDP-mCherry. Magnification,×400. TF, tissue factor; hTF, human tissue factor.