Evaluating Exosome Effects on Dendritic Cell Apoptosis

BMDCs were generated from bone marrow cells isolated from C57BL/6 mice, as previously described [17 (link)]. Day 8 immature BMDCs were incubated with lipopolysaccharide (LPS, 100 ng/mL; Invitrogen, San Diego, CA, USA) or exosomes (5 and 25 μg/mL) isolated from non-irradiated (N-exo) and gamma-irradiated cancer cells (G-exo) for 18 h in a humidified incubator containing 5% CO₂ at 37 °C. Cells were then harvested and stained with Annexin V and PI (BD Bioscience) according to the systematic instructions provided by the manufacturer. Samples (stained cells) were analyzed using a FACS instrument (MACSQuant VYB; Miltenyi Biotec, San Diego, CA, USA).