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Figure 3 Mobilization of putative vascular progenitor cells by granulocyte-colony stimulating factor (G-CSF) and in vitro differentiation to vascular smooth muscle cell (SMC) or endothelial cell (EC). Four quadrant analysis of antigen expression in circulating peripheral blood mononuclear cells after 6 days of G-CSF or placebo injection, and serial observation of cell culture with vascular endothelial growth factor (VEGF) or platelet-derived growth factor (PDGF). Endothelial lineage cells were identified by CD31 and VE-Cadherin (VE-CAD) in FACS analysis and PE-labeled-CD31 immunocytochemical staining (red color), and vascular smooth muscle lineage cells were identified by alpha-smooth muscle actin (SMA) in FACS analysis and FITC-alpha-SMA immunostaining (green color). Both endothelial and smooth muscle lineage cells increased with G-CSF treatment in peripheral blood. During culture of peripheral blood mononuclear cells, endothelial lineage cells increased preferentially with VEGF. In contrast, PDGF induced a part of the cells to differentiate into smooth muscle lineage in addition to double positive (CD31+/alpha-SMA+) or endothelial lineage cells.