Mixed culture of moLCs and CD1a+Langerin cells (50,000 cells per well) were seeded in CellCarrier Ultra plates (PerkinElmer, Waltham, MA) supplemented with 50 ng/ml GM-CSF. They were stained with phycoerythrin-conjugated Langerin (clone MB22-9F5, Miltenyi), Hoechst33342 fluorescence dye (Thermo Fischer Scientific), and 1 μM lysotracker (Thermo Fischer Scientific). A647-labeled Langerin ligand‒targeted or non-targeted liposomes were added for imaging at a lipid concentration of 16 μM. Live cell imaging was performed with a ×63 lens, and pictures were acquired every 5 minutes over 1 hour at 37 °C on the Operetta High-Content Imaging System (PerkinElmer). Samples were analyzed using the Operetta Harmony Software (PerkinElmer).

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