Exosome-Mediated Drug Delivery Protocol

HEK293T exosomes (10¹¹ particles/ml) were mixed with tazemetostat (1µM) (HY-130803, MedChemExpress) and enzalutamide (10 µM) (MDV-3100, Biovision) in 1ml of PBS followed by sonication. Sonication settings used were 20% amplitude, 500V voltage and 2kHz frequency. 6 cycles of sonication were used with 30 s ‘pulse on’ and 2 min of ‘pulse off’. After sonication, exosome samples were incubated at 37 °C for 1 h for recovery. Subsequently, the exosomes were purified using the Total exosome isolation kit (Invitrogen, cat no. 4478359) according to the manufacturer's instructions. Finally, the purified exosomes were visualized and quantified using a Nanosight LM10 instrument (Malvern Instruments).