Figure 3: Enumeration of circulating EPC-EC. Example of gating strategy of EPC-EC by FACS. A representative FACS demonstrates enumeration of EPC and EC analyzed fresh and after cryopreservation. A. PBMC was assessed as viable by negative staining for DAPI. B. CD45+ cells were excluded and the rest of the cell population was further gated for CD34+ cells. C. CD34+ cells were gated for CD133 and VEGFR2 for enumeration of double and triple positive subsets of EPC. D. CD 45+ cells were excluded and live cells were further gated for CD146+. E. CD 146+ cells were further gated for CD34 and VEGFR2/ (KDR) for enumeration of double and triple positive subsets of EC.