Figure 2

Analysis of rec41D3-induced pSn endocytosis and analysis of colocalization between internalized antibody and endo/lysosomal compartments. (A) Confocal microscopical analysis of mAb 41D3- and rec41D3-induced pSn internalization in primary macrophages. Cells were incubated for the indicated time periods with either antibody at 37°C, fixed, permeabilized and stained with FITC-labelled goat-anti-mouse IgG. Images represent a single confocal z-section through the middle of the cell. (B) Analysis of colocalization between internalized mAb 41D3 or rec41D3 (green) and early endosomes (EEA1), late endosomes (CI-M6P) or lysosomes (Lamp1) (red) in primary macrophages. Colocalization was calculated from confocal z-sections of 25 randomly selected cells of 2 independent experiments at the indicated time points. Data represent the means ± standard deviations. Representative images of macrophages that were incubated for 60 minutes at 37°C with either antibody are shown as overlays of the green and red signal with a yellow colour indicating colocalization. Scale bar: 5 μm.