Figure 4. Late endosomal/lysosomal trafficking analysis in HEK293-hFcRn-GFP cells.

HEK293-hFcRn-GFP cells were incubated with 50 nM AF647-labeled FcRn antagonist or IgG1-WT (control) for 3 hours. Following incubation, cells were fixed and permeabilized, and detection of late endosomes/lysosomes was carried out using anti–LAMP-1 antibody followed by AF555-labeled goat anti-mouse IgG conjugate. Yellow arrowheads indicate the detection of hFcRn-GFP and AF647 antagonists in anti–LAMP-1–positive compartments. Images for the AF555 channel were adjusted for visualization. Data are representative of 2 independent experiments, each consisting of at least 2 dishes per condition, and at least 6 images for each dish. AF555, AF647, and GFP are pseudocolored red, blue, and green, respectively. Each image represents part of a single cell. Scale bars = 2 μm.