Uptake of 2B3-101 in human brain endothelial hCMEC/D3 cells.

<p>(A) Representative graph (mean ± SEM, n = 3 per time point) of the time-dependent experiment at 37°C showed a higher uptake of 2B3-101 compared to pegylated liposomal doxorubicin (PEG-lipo-DOX), both at a concentration of 450 µg/ml HSPC. (B) Representative graph of the concentration-dependent experiments at 4 and 37°C (mean ± SEM, n = 3 per concentration). Cells incubated at 4°C (dotted lines) show a higher fluorescence when treated with pegylated liposomal doxorubicin than with 2B3-101. However, at 37°C (full lines), 2B3-101 incubation resulted in a stronger fluorescence than pegylated liposomal doxorubicin. (C) The percentage of bound 2B3-101 being internalized [(MFI@37°C−MFI@4°C)/MFI@4°C] is increasing in a concentration-dependent manner for 2B3-101, while pegylated liposomal doxorubicin mainly remains bound to the cells. The overall mean ± SEM from 3 independent experiments (each n = 3) was used in a paired t-test. *p<0.05 2B3-101 vs. PEG-lipo-DOX, both at 450 µg/ml HSPC and 2B3-101 at 450 µg/ml vs. 50 µg/ml.