Plasma Exosome Isolation Protocol

The collected plasma was thawed at 37 °C and then centrifuged at 3000 g for 15 min to remove cell debris. Aspirated supernatant was diluted sevenfold with PBS and centrifuged at 13 000 g for 30 min [24 (link)]. Large particles were removed using 0.22‐μm filters. The collected supernatant was then ultra‐centrifuged at 100 000 g, 4 °C for 2 h (CP100NX; Hitachi, Brea, CA, USA). The pellet containing exosomes was re‐suspended in PBS and ultra‐centrifuged again at 100 000 g 4 °C for 2 h. The isolated exosomes were re‐suspended in 100 µL PBS after PBS washing for further analysis.