Fig. 6: The scheme showing the mass spectrometry (MS)-based trans-SILAC approach performed to identify proteins that are present within stroma-derived vesicles.

Donor stromal cells were cultured in media containing heavy isotopologues of Lys and Arg for 9 days to allow complete incorporation of these amino acids into their proteome. They were stained with DiD for cellular vesicles and subjected to co-culture with acceptor CML cells for 24 h. Afterwards two subpopulations of CML cells were sorted using FACS: cells which received (DiD+) or did not receive (DiD-) cellular vesicles from donor stromal cells. In both samples, heavy proteins transferred from donor stromal cells, were identified. Only proteins present exclusively in the DiD+ sample were considered to have been transferred along with the studied vesicles