4.12. Analysis of miR-153 Secretion in Exosomes and Microvesicles

In order to quantify the miR-153 in exosome and microvesicles, we first collected the 48 h conditional medium from 70% confluent cells, both for PC3 and DU145. Then, the exosomes and microvesicles were isolated by differential ultracentrifugation, as described above. The RNA was extracted, and the analysis of miR-153 was performed by RT-qPCR using the MystiCq microRNA cDNA synthesis kit (MIRRT) (Sigma Aldrich, Burlington, MA, USA). The level of expression of miR-153 in each sample was normalized on an internal control of the kit. PC3/DU145 exosomal and microvesicle miR-153 expression was compared to that of cellular PC3 or DU145.