Fig. 5.

Kinetics of antibody internalization and subcellular localization. DU145 cells were incubated with three different IgGs (HCA-F1, HCA-M1, or HCA-S1) at 10 μg/ml for 15 min at 4 °C and then chased with complete DMEM/10% FBS for indicated time periods. Cells were then fixed, permeabilized, and immunolabeled against human IgG (green) and A, early endosomes (EEA1, red) or B, lysosomes (LAMP1, red). Nuclei were stained with Hoechst 33342 (blue). Scale bar: 20 μm. Pearson's correlation coefficients between immunolabeled C, EEA1 or D, LAMP1 and immunolabeled IgG were averaged from a minimum of 30 cells. Error bars denote S.E. of n = 3.