The hermaphrodite strain N2 (Bristol), fed with Escherichia coli OP50, was maintained and synchronized. Young adults were thoroughly washed before incubations for 5 h or 24 h in M9 buffer. Between 35,500 and 158,000 worms were used for each experiment. After incuabtion, supernatants were processed by differential centrigugation/ultracentrifugation to pellet extracellular vesicles and characterization of the protein contents by mass spectrometry.