Figure 1.

Donor EV in recipient kidneys. (A and B) ICAM1 and CD63 protein levels in normoxic EV (EXO) and IPC EV (IPC EXO), measured on western blot, n=5. ICAM1 was higher (P=0.001) and CD63 lower (P<0.01) in IPC EXO. (C) Electron microscopy showed IPC EXO and EXO (not shown) averaged <200 nM diameter. (D) RT-PCR was used to identify the SRY male determining gene in EV (EXO) from male donor cells (positive control), and also in kidneys of female EV recipients (IPC EXO and EXO), but not in female kidneys of rats injected with saline (saline). Additional SRY-positive controls were amplified from normal male rat kidneys. (E) Representative image of three rat kidneys used to localize IPC EV labeled with red EXO-Glow. A 10% fraction of IPC EV was labeled, remixed with the unlabeled 90%, injected, and visualized by fluorescent microscopy in 50-μM vibratome sections. (F) RT-PCR was used to identify the murine SAA1 gene amplified in donor EV derived from cells transfected with SAA1 (EXO), and also in recipient kidneys of rats injected with EXO and IPC EXO (EXO and IPC EXO), but not in kidneys of rats injected with saline (saline). The negative and positive controls were amplifications of pCDNA1 and SAA1-pCDNA1 plasmids used for transfection. Ctrl, control; MW, molecular weight; pos, positive.