Figure 2.

In vitro characterization of the antagonistic uPAR antibodies. (A) Internalization of 2G10, 3C6 and 1A8 labeled with ¹¹¹In at 37^oC. MDA-MB-231 expressing uPAR and MDA-MB-231 (uPAR-) cells with expression knocked out were incubated with 10 nM of radiolabeled antibody at the indicated time points and were washed and treated with an acidic buffer to remove non-covalently bound and non-internalized antibody. Each time point was performed in triplicate. (B) Internalization of ¹¹¹In-2G10 at the 120 min time point by the drug-resistant and parental cell lines cells lines. Blocking was performed by adding 1 µM of cold 2G10 IgG or 1 µM of 2G10 prior to addition of radiolabeled antibody. (C) Staining of different breast cancer cell lines with FITC labeled 2G10 as analyzed by flow cytometry.